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Gene Sequences (gene + sequence)
Kinds of Gene Sequences Terms modified by Gene Sequences Selected AbstractsSYSTEMATICS OF THE HILDENBRANDIALES (RHODOPHYTA): GENE SEQUENCE AND MORPHOMETRIC ANALYSES OF GLOBAL COLLECTIONS,JOURNAL OF PHYCOLOGY, Issue 2 2003Alison R. Sherwood Fifty-seven collections of marine and freshwater Hildenbrandia from North America, South America, Europe, and Africa were compared with 21 type and historically important specimens using multivariate morphometrics. Additionally, phylogenetic analyses of 48 specimens of Hildenbrandia and two specimens of Apophlaea were carried out based on sequences of the rbcL chloroplast gene and the nuclear 18S rRNA gene. Morphometric analyses based on vegetative cell and filament dimensions distinguished two groups of freshwater Hildenbrandia specimens, the first corresponding to those collections from North America and the Philippines and the second to those from Europe and the Canary Islands. The first group had smaller mean cell and filament dimensions (cells 4.0 × 4.4 ,m, filaments 46.5 ,m) and corresponded to H. angolensis, whereas the second group had larger mean dimensions (cells 5.8 × 6.6 ,m, filaments 55.3 ,m) and represented H. rivularis. Marine specimens were morphometrically distinguishable into two groups based on tetrasporangial division pattern as well as other thallus characters. However, measurements and character determinations of some type specimens differed greatly from the original descriptions, and thus further work to determine the stability of these characters is required. Phylogenetic reconstruction based on the 18S rRNA gene and rbcL gene sequence data generally demonstrated separation of the marine and freshwater forms of Hildenbrandia, with some marine taxa forming monophyletic groups (e.g. H. lecannellieri and H. occidentalis) and others forming paraphyletic groups (e.g. H. rubra). The two specimens of Apophlaea formed a monophyletic group within the paraphyletic genus Hildenbrandia. [source] UREASE GENE SEQUENCES FROM ALGAE AND HETEROTROPHIC BACTERIA IN AXENIC AND NONAXENIC PHYTOPLANKTON CULTURES,JOURNAL OF PHYCOLOGY, Issue 3 2009Kristopher M. Baker While urea has long been recognized as an important form of nitrogen in planktonic ecosystems, very little is known about how many or which phytoplankton and bacteria can use urea as a nitrogen source. We developed a method, targeting the gene encoding urease, for the direct detection and identification of ureolytic organisms and tested it on seven axenic phytoplankton cultures (three diatoms, two prymnesiophytes, a eustigmatophyte, and a pelagophyte) and on three nonaxenic Aureococcus anophagefferens Hargraves et Sieburth cultures (CCMP1784 and two CCMP1708 cultures from different laboratories). The urease amplicon sequences from axenic phytoplankton cultures were consistent with genomic data in the three species for which both were available. Seven of 12 phytoplankton species have one or more introns in the amplified region of their urease gene(s). The 63 urease amplicons that were cloned and sequenced from nonaxenic A. anophagefferens cultures grouped into 17 distinct sequence types. Eleven types were related to ,-Proteobacteria, including three types likely belonging to the genus Roseovarius. Four types were related to ,-Proteobacteria, including two likely belonging to the genus Marinobacter, and two types were related to ,-Proteobacteria. Terminal restriction fragment length polymorphism (TRFLP) analyses suggested that the sequenced amplicons represented approximately half of the diversity of bacterial urease genes present in the nonaxenic cultures. While many of the bacterial urease sequence types were apparently lab- or culture-specific, others were found in all three nonaxenic cultures, suggesting the possibility of specific relationships between these bacteria and A. anophagefferens. [source] PHYLOGENY OF THE DASYCLADALES (CHLOROPHYTA, ULVOPHYCEAE) BASED ON ANALYSES OF RUBISCO LARGE SUBUNIT (rbcL) GENE SEQUENCES,JOURNAL OF PHYCOLOGY, Issue 4 2003Frederick W. Zechman The phylogeny of the green algal Order Dasycladales was inferred by maximum parsimony and Bayesian analyses of chloroplast-encoded rbcL sequence data. Bayesian analysis suggested that the tribe Acetabularieae is monophyletic but that some genera within the tribe, such as Acetabularia Lamouroux and Polyphysa Lamouroux, are not. Bayesian analysis placed Halicoryne Harvey as the sister group of the Acetabularieae, a result consistent with limited fossil evidence and monophyly of the family Acetabulariaceae but was not supported by significant posterior probability. Bayesian analysis further suggested that the family Dasycladaceae is a paraphyletic assemblage at the base of the Dasycladales radiation, casting doubt on the current family-level classification. The genus Cymopolia Lamouroux was inferred to be the basal-most dasycladalean genus, which is also consistent with limited fossil evidence. Unweighted parsimony analyses provided similar results but primarily differed by the sister relationship between Halicoryne Lamouroux and Bornetella Munier-Chalmas, thus supporting the monophyly of neither the families Acetabulariaceae nor Dasycladaceae. This result, however, was supported by low bootstrap values. Low transition-to-transversion ratios, potential loss of phylogenetic signal in third codon positions, and the 550 million year old Dasycladalean lineage suggest that dasyclad rbcL sequences may be saturated due to deep time divergences. Such factors may have contributed to inaccurate reconstruction of phylogeny, particularly with respect to potential inconsistency of parsimony analyses. Regardless, strongly negative g1 values were obtained in analyses including all codon positions, indicating the presence of considerable phylogenetic signal in dasyclad rbcL sequence data. Morphological features relevant to the separation of taxa within the Dasycladales and the possible effects of extinction on phylogeny reconstruction are discussed relative to the inferred phylogenies. [source] MOLECULAR SYSTEMATICS OF RIVER DOLPHINS INFERRED FROM COMPLETE MITOCHONDRIAL CYTOCHROME- B GENE SEQUENCESMARINE MAMMAL SCIENCE, Issue 1 2002Guang Yang Abstract 1,140 bp of the complete mitochondrial cytochrome- b gene sequences of baiji (Lipotes vexillifer), franciscana (Pontoporia blainvillei), and Ganges river dolphin (Platanista gangetica gangetica) were determined to address the systematic position and phylogeny of extant river dolphins with combination of homologous sequences of other cetaceans. The neighbor-joining (NJ), maximum parsimony (MP), and maximum likelihood (ML) phylogenetic analyses all identified the river dolphins into three lineages, i. e., Platanista, Lipotes, and Inia+Pontoporia. The Lipotes did not have sister relationship with either Platanista or Inia+Pontoporia, which strongly supported the referral of Lipotes to a separate family, i. e., Lipotidae. There were very high sequence divergences between all river dolphin genera, suggesting a relatively longer period of separation time than those among other odontocete families. [source] Study of the Cytochrome b Gene Sequence in Populations of TaiwanJOURNAL OF FORENSIC SCIENCES, Issue 1 2010Hsiao-Lin Hwa M.D., Ph.D. Abstract:, The cytochrome b gene (MTCYB) has been widely used in taxonomic research. In this study, the sequence polymorphism of the MTCYB gene was determined in 417 subjects of eight populations living in Taiwan (Taiwanese Han, indigenous Taiwanese, Tao, mainland Chinese, Filipino, Thai, Vietnamese, and Caucasian). Sequence variation from the revised Cambridge Reference Sequence and genetic distance between these populations were analyzed. There were 108 variable positions with a total of 99 haplotypes. Population-specific positions of MTCYB gene were noted in Tao and Caucasian populations. There were statistically significant differences of genetic distance between Taiwanese Han and Caucasian, between Taiwanese Han and Tao, and between Taiwanese Han and Filipino. A phylogenetic tree presents the genetic distances between these populations. In conclusion, there are sufficient sequence polymorphisms of the MTCYB gene in individuals of different populations, which may be used in the analyses of human ethnic groups in forensic casework. [source] Phylogenetic Analysis of Complete rRNA Gene Sequence of Nosema philosamiae Isolated from the Lepidopteran Philosamia cynthia riciniTHE JOURNAL OF EUKARYOTIC MICROBIOLOGY, Issue 3 2010FENG ZHU ABSTRACT. The microsporidian Nosema philosamiae is a pathogen that infects the eri-silkworm Philosamia cynthia ricini. The complete sequence of rRNA gene (4,314 bp) was obtained by polymerase chain reaction amplification with specific primers and sequencing. The sequence analysis showed that the organization of the rRNA of N. philosamiae was similar to the pattern of Nosema bombycis. Phylogenetic analysis of rRNA gene sequences revealed that N. philosamiae had a close relationship with other Nosema species, confirming that N. philosamiae is correctly assigned to the genus Nosema. [source] Stoeckeria algicida n. gen., n. sp. (Dinophyceae) from the Coastal Waters off Southern Korea: Morphology and Small Subunit Ribosomal DNA Gene SequenceTHE JOURNAL OF EUKARYOTIC MICROBIOLOGY, Issue 4 2005HAE JIN JEONG Abstract. This paper presents a new description of the morphology of the planktonic dinoflagellate Stoeckeria algicida n. gen., n. sp. and a report of the sequence of the small subunit rDNA (SS rDNA) from cultured cells. The vegetative biflagellated cell, gametes, triflagellated planozygotes, and cyst stages of this heterotrophic species were observed in cultures. The vegetative biflagellated cells are oval, with the cell length being considerably larger than the cell width. The ranges (and mean, n=60) of cell length and width of live biflagellated cells satiated with the raphidophyte Heterosigma akashiwo were 14.4,20.8 ,m (16.8) and 10.0,17.4 ,m (12.9), respectively, while those of biflagellated cells starved for 3 d (n=60) were 7.3,15.9 ,m (11.6) and 2.7,12.2 ,m (7.3), respectively. Thin plates of the vegetative biflagellated cells were arranged in a Kofoidian series of Po, cp, X, 4,, 2a, 7,, 6c, 6s, 5,,, 0 (p), and 2,,. When properly aligned, the sequence of the SS rDNA of the biflagellated cells of S. algicida (GenBank Accession no. AJ841809) was 3% different from that of a dinoflagellate from Shepherd's Crook and 4% different from that of Cryptoperidiniopsoid sp. brodyi, Pfiesteria spp., or Pfiesteria -like species. In a maximum-likelihood-distance phylogenetic tree generated using the SS rDNA sequences, Pfiesteria spp., Pfiesteria- like species, and a dinoflagellate from Shepherd's Crook were closest to S. algicida, but these dinoflagellates were clearly divergent with S. algicida. Based on morphological and genealogical analyses, we suggest that this is a new species in a new genus. [source] Taxonomic Redescriptions of Two Ciliates, Protogastrostyla pulchra n. g., n. comb. and Hemigastrostyla enigmatica (Ciliophora: Spirotrichea, Stichotrichia), with Phylogenetic Analyses Based on 18S and 28S rRNA Gene SequencesTHE JOURNAL OF EUKARYOTIC MICROBIOLOGY, Issue 6 2007JUN GONG ABSTRACT. The morphology and infraciliature of two stichotrichid ciliates, Gastrostyla pulchra(Perejaslawzewa 1886) Kahl, 1932 and Hemigastrostyla enigmatica(Dragesco and Dragesco-Kernéis 1986) Song & Wilbert, 1997, collected from marine and brackish sediments, were investigated by using living observations and protargol impregnations. Both 18S and 28S rRNA genes of these two species were sequenced. The 18S rDNA show high similarities (98.4%,99.7%) among populations of each species. There is about 94% similarity in 18S rDNA genes between G. pulchra and Gastrostyla steinii, the type species of the genus, which has been confirmed to be an oxytrichid by previous studies. In the phylogenetic trees of 18S, 28S, and combined 18S and 28S rDNA, both G. pulchra and H. enigmatica are consistently placed outside the well-established oxytrichid clade. Based on our analyses and previous ontogenetic data, we conclude that these two species may represent some lower groups in the subclass Stichotrichia, and that G. pulchra should represent a new genus, Protogastrostyla n. g. This new genus, which is morphologically similar to Gastrostyla, differs in its morphogenesis: the apical part of the old AZM is retained combining with the newly built membranelles that develop from the proter's oral primordium; the primary primordia of the dorsal kinety; and marginal primordia commence de novo without a definite contribution from the old structure. [source] Reevaluation of the Phylogenetic Relationship between Mobilid and Sessilid Peritrichs (Ciliophora, Oligohymenophorea) Based on Small Subunit rRNA Genes SequencesTHE JOURNAL OF EUKARYOTIC MICROBIOLOGY, Issue 5 2006YING-CHUN GONG ABSTRACT. Based on morphological characters, peritrich ciliates (Class Olygohymenophorea, Subclass Peritrichia) have been subdivided into the Orders Sessilida and Mobilida. Molecular phylogenetic studies on peritrichs have been restricted to members of the Order Sessilida. In order to shed more light into the evolutionary relationships within peritrichs, the complete small subunit rRNA (SSU rRNA) sequences of four mobilid species, Trichodina nobilis, Trichodina heterodentata, Trichodina reticulata, and Trichodinella myakkae were used to construct phylogenetic trees using maximum parsimony, neighbor joining, and Bayesian analyses. Whatever phylogenetic method used, the peritrichs did not constitute a monophyletic group: mobilid and sessilid species did not cluster together. Similarity in morphology but difference in molecular data led us to suggest that the oral structures of peritrichs are the result of evolutionary convergence. In addition, Trichodina reticulata, a Trichodina species with granules in the center of the adhesive disc, branched separately from its congeners, Trichodina nobilis and Trichodina heterodentata, trichodinids without such granules. This indicates that granules in the adhesive disc might be a phylogenetic character of high importance within the Family Trichodinidae. [source] Real-time primer design for DNA chipsCONCURRENCY AND COMPUTATION: PRACTICE & EXPERIENCE, Issue 9 2004H. Simmler Abstract The design of PCR or DNA chip experiments is a time-consuming process where bioinformatics is extensively used. The selection of the primers, which are immobilized on the DNA chip, requires a complex algorithm. Based on several parameters an optimized set of primers is automatically determined for a given gene sequence. This paper describes a parallel architecture which performs the optimization of the primer selection on a hardware accelerator. In contrast to the pure software approach, the parallel architecture gains a speedup of factor 500 using a PCI-based hardware accelerator. This approach allows an optimization of a specified primer set in real time. Copyright © 2004 John Wiley & Sons, Ltd. [source] Isolation and gene quantification of heterotrophic N2 -fixing bacterioplankton in the Baltic SeaENVIRONMENTAL MICROBIOLOGY, Issue 1 2007Kjärstin H. Boström Summary Cyanobacteria are regarded as the main N2 -fixing organisms in marine waters. However, recent clone libraries from various oceans show a wide distribution of the dinitrogenase reductase gene (nifH) originating from heterotrophic bacterioplankton. We isolated heterotrophic N2 -fixing bacteria from Baltic Sea bacterioplankton using low-nitrogen plates and semi-solid diazotroph medium (SSDM) tubes. Isolates were analysed for the nitrogenase (nifH) gene and active N2 fixation by nested polymerase chain reaction (PCR) and acetylene reduction respectively. A primer-probe set targeting the nifH gene from a , - proteobacterial isolate, 97% 16S rDNA similarity to Pseudomonas stutzeri, was designed for measuring in situ dynamics using quantitative real-time PCR. This nifH gene sequence was detected at two of 11 stations in a Baltic Proper transect at abundances of 3 × 104 and 0.8 × 103 copies per litre seawater respectively. Oxygen requirements of isolates were examined by cultivation in SSDM tubes where oxygen gradients were determined with microelectrodes. Growth, and thereby N2 fixation, was observed as horizontal bands formed at oxygen levels of 0,6% air saturation. The apparent microaerophilic or facultative anaerobic nature of the isolates explains why the SSDM approach is the most appropriate isolation method. Our study illustrates how combined isolation, functional analyses and in situ quantification yielded insights into the oxygen requirements of heterotrophic N2 -fixing bacterioplankton isolates, which were confirmed to be present in situ. [source] Genetic and functional properties of uncultivated thermophilic crenarchaeotes from a subsurface gold mine as revealed by analysis of genome fragmentsENVIRONMENTAL MICROBIOLOGY, Issue 12 2005Takuro Nunoura Summary Within a phylum Crenarchaeota, only some members of the hyperthermophilic class Thermoprotei, have been cultivated and characterized. In this study, we have constructed a metagenomic library from a microbial mat formation in a subsurface hot water stream of the Hishikari gold mine, Japan, and sequenced genome fragments of two different phylogroups of uncultivated thermophilic Crenarchaeota: (i) hot water crenarchaeotic group (HWCG) I (41.2 kb), and (ii) HWCG III (49.3 kb). The genome fragment of HWCG I contained a 16S rRNA gene, two tRNA genes and 35 genes encoding proteins but no 23S rRNA gene. Among the genes encoding proteins, several genes for putative aerobic-type carbon monoxide dehydrogenase represented a potential clue with regard to the yet unknown metabolism of HWCG I Archaea. The genome fragment of HWCG III contained a 16S/23S rRNA operon and 44 genes encoding proteins. In the 23S rRNA gene, we detected a homing-endonuclease encoding a group I intron similar to those detected in hyperthermophilic Crenarchaeota and Bacteria, as well as eukaryotic organelles. The reconstructed phylogenetic tree based on the 23S rRNA gene sequence reinforced the intermediate phylogenetic affiliation of HWCG III bridging the hyperthermophilic and non-thermophilic uncultivated Crenarchaeota. [source] Isolation and properties of methanesulfonate-degrading Afipia felis from Antarctica and comparison with other strains of A. felisENVIRONMENTAL MICROBIOLOGY, Issue 1 2005S. Azra Moosvi Summary Three novel strains of methylotrophic Afipia felis were isolated from several locations on Signy Island, Antarctica, and a fourth from estuary sediment from the River Douro, Portugal. They were identified as strains of the ,-2 proteobacterium A. felis by 16S rRNA gene sequence, analysis., Two, strains, tested, were, shown to contain the fdxA gene, diagnostic for A. felis. All strains grew with methanesulfonate (and two strains with dimethylsulfone) as sole carbon substrate. Growth on methanesulfonate required methanesulfonate monooxygenase (MSAMO), using NADH as the reductant and stimulated by reduced flavin nucleotides and Fe(II). Polymerase chain reaction amplification of DNA from an Antarctic strain showed a typical msmA gene for the ,-hydroxylase of MSAMO, and both Antarctic and Portuguese strains contained mxaF, the methanol dehydrogenase large subunit gene. This is the first report of methanesulfonate-degrading bacteria from the Antarctic and of methylotrophy in Afipia, and the first description of any bacterium able to use both methanesulfonate and dimethylsulfone. In contrast, the type strain of A. felis DSM 7326T was not methylotrophic, but grew in defined mineral medium with a wide range of single simple organic substrates. Free-living Afipia strains occurring widely in the natural environment may be significant as methylotrophs, degrading C1 -sulfur compounds, including the recalcitrant organosulfur compound methanesulfonate. [source] Early diagnosis of rhinocerebral mucormycosis by cerebrospinal fluid analysis and determination of 16s rRNA gene sequenceEUROPEAN JOURNAL OF NEUROLOGY, Issue 9 2007D. Bengel A 40-year-old diabetic woman was diagnosed with rhinocerebral mucormycosis. Cerebral mucormycosis is an acute life-threatening disease, which is caused by fungi of the class Phycomycetae. Clinical suspicion and detection of the fungal hyphae in cerebrospinal fluid (CSF) led to early diagnosis, subsequently confirmed by immunohistochemistry and molecular analysis of fungal RNA. Early infiltration of the infectious agent into the central nervous system resulted in septic thrombosis of the cavernous sinus, mycotic meningoencephalitis, brain infarctions as well as intracerebral and subarachnoidal hemorrhages. Despite immediate high-dose antimycotic treatment, surgical debridement of necrotic tissue, and control of diabetes as a predisposing factor, the woman died 2 weeks after admission. Although fungal organisms are rarely detectable in CSF specimens from patients with mycotic infections of the central nervous system, comprehensive CSF examination is beneficial in the diagnosis of rhinocerebral mucormycosis. Furthermore, a concerted team approach, systemic antifungal agents and early surgical intervention seem to be crucial for preventing rapid disease progression. [source] Lessons from leeches: a call for DNA barcoding in the labEVOLUTION AND DEVELOPMENT, Issue 6 2006Alexandra E. Bely SUMMARY Many evolution of development labs study organisms that must be periodically collected from the wild. Whenever this is the case, there is the risk that different field collections will recover genetically different strains or cryptic species. Ignoring this potential for genetic variation may introduce an uncontrolled source of experimental variability, leading to confusion or misinterpretation of the results. Leeches in the genus Helobdella have been a workhorse of annelid developmental biology for 30 years. Nearly all early Helobdella research was based on a single isolate, but in recent years isolates from multiple field collections and multiple sites across the country have been used. To assess the genetic distinctness of different isolates, we obtained specimens from most Helobdella laboratory cultures currently or recently in use and from some of their source field sites. From these samples, we sequenced part of the mitochondrial gene cytochrome oxidase I (COI). Sequence divergences and phylogenetic analyses reveal that, collectively, the Helobdella development community has worked on five distinct species from two major clades. Morphologically similar isolates that were thought to represent the same species (H. robusta) actually represent three species, two of which coexist at the same locality. Another isolate represents part of a species complex (the "H. triserialis" complex), and yet another is an invasive species (H. europaea). We caution researchers similarly working on multiple wild-collected isolates to preserve voucher specimens and to obtain from these a molecular "barcode," such as a COI gene sequence, to reveal genetic variation in animals used for research. [source] Genetic differentiation of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) biotype Q based on mitochondrial DNA markersINSECT SCIENCE, Issue 2 2008Dong Chu Abstract In the present study, genetic differentiation of Bemisia tabaci (Gennadius) biotype Q was analyzed based on mitochondrial cytochrome oxidase I (mt COI) gene sequence. The results showed that B. tabaci biotype Q could be separated into two subclades, which were labeled as subclades Q1 and Q2. Subclade Q1 was probably indigenous to the regions around the Mediterranean area and subclade Q2 to Israel or Cyprus. It was because B. tabaci was composed of several genetically distinct groups with a strong geographical association between more closely related biotypes. Not all of the B. tabaci biotype Q in the non-Mediterranean countries come from the same regions. Until now, all B. tabaci biotype Q in China were grouped into subclade Q1. The B. tabaci biotype Q introduced into the US included both subclades Q1 and Q2. The genetic structure analysis showed higher genetic variation of subclade Q1 than that of subclade Q2. [source] Frequency distribution of a Cys430Ser polymorphism in peroxisome proliferator-activated receptor-gamma coactivator-1 (PPARGC1) gene sequence in Chinese and Western pig breedsJOURNAL OF ANIMAL BREEDING AND GENETICS, Issue 1 2005T. Kunej Summary Identification of major genes, that genetically impact fat tissue formation is important for successful selection of lean animals with good meat quality. Because of its central role in fat cell differentiation and muscle fibre type determination, PPARGC1 is a potential candidate gene affecting fattening traits and pig meat quality. In this study, a T/A substitution at position 1378 (GenBank accession no. AY346131) in the porcine PPARGC1 gene causing a Cys430Ser amino acid substitution at position 430 was genotyped on a total of 239 animals, including 101 from seven Chinese and 138 from six Western pig breeds. Bayesian analysis revealed that the mean frequency of allele T (Cys) was 92.64 ± 4.82% in Chinese pigs, and 45.99 ± 4.13% in Western pigs. The 95% interval of the posterior mean frequency of allele T was 0.82,1.00 in Chinese pigs and 0.38,0.54 in Western pigs, indicating these two groups of pigs diverged at this locus during genetic evolution of the breed. Because marked differences in fat and lean tissue deposition exist between Western and Chinese pig breeds, this Cys430Ser exchange in the PPARGC1 gene deserves further evaluation to determine its phenotypic effect on fattening and carcass traits in commercial pig populations. [source] Production of enterolysin A by rumen Enterococcus faecalis strain and occurrence of enlA homologues among ruminal Gram-positive cocciJOURNAL OF APPLIED MICROBIOLOGY, Issue 2 2007K. Nigutova Abstract Aims:, Purification and partial characterization of an extracellular bacteriocin produced by the ruminal isolate Enterococcus faecalis II/1 and determine the frequency of occurrence of enterolysin A structural gene within the ruminal cocci. Methods and Results:, Bacteriocin produced by E. faecalis II/1 was purified to homogeneity. Purified bacteriocin exhibited a single band on sodium dodecylsulphate polyacrylamide gel electrophoresis with an apparent molecular weight of about 35 kDa. The amino acid sequence of the first 30 amino acids of purified bacteriocin was identical with the enterolysin A sequence. The DNA sequence of the nearly complete E. faecalis II/1 bacteriocin structural gene was identical to the enterolysin A gene sequence, confirming that this bacteriocin is identical to enterolysin A, a cell wall-degrading bacteriocin from E. faecalis LMG 2333. Enterolysin A structural genes were detected in approximately one-sixth of the Gram-positive ruminal cocci examined by PCR using primers targeting the enterolysin A structural gene. Conclusions:, Bacteriocin produced by E. faecalis II/1 is identical to enterolysin A. Enterolysin A structural gene homologues are frequently encountered in rumen enterococcal and streptococcal bacterial strains. Significance and Impact of the study:, This is the first evidence of a large heat-labile bacteriocin produced by rumen E. faecalis strain, enlarging the number and types of known anti-bacterial proteins produced by rumen bacteria. [source] Isolation and characterization of a bacterial strain of the genus Ochrobactrum with methyl parathion mineralizing activityJOURNAL OF APPLIED MICROBIOLOGY, Issue 5 2006X.-H. Qiu Abstract Aims:, To isolate and characterize a methyl parathion (MP)-mineralizing bacterium, and to elucidate the degradative pathway of MP and localize the responsible degrading genes. Methods and Results:, A bacterial strain, designated B2, capable of mineralizing MP was isolated from the MP-polluted soil. Analysis of the 16S rRNA gene sequence and phenotypic analysis suggested that strain B2 had a close relationship with Ochrobactrum anthropi. B2 could totally degrade MP and four metabolites [p -nitrophenol (PNP), 4-nitrocatechol (4-NC), 1,2,4-benzenetriol (BT) and hydroquinone (HQ)] were identified by HPLC and gas chromatography-mass spectrometry analyses. Plasmid curing of strain B2 resulted in the loss of ability of B2 to degrade PNP, but not the ability to hydrolyse MP. Conclusions:,Ochrobactrum sp. B2 can mineralize MP rapidly via PNP, 4-NC, BT and HQ pathway. B2 harbours a plasmid encoding the ability to degrade PNP, while MP-hydrolysing activity is encoded on the bacterial chromosome. Significance and Impact of the Study:, This new bacterial strain (B2) capable of mineralizing MP will be useful in a pure-culture remediation process of organophosphate pesticides and their metabolites such as nitroaromatics. [source] Physiological characterization of Mycobacterium sp. strain 1B isolated from a bacterial culture able to degrade high-molecular-weight polycyclic aromatic hydrocarbonsJOURNAL OF APPLIED MICROBIOLOGY, Issue 2 2004C.E. Dandie Abstract Aim:, The aim of this study was to further characterize a bacterial culture (VUN 10,010) capable of benzo[a]pyrene cometabolism. Methods and Results:, The bacterial culture, previously characterized as a pure culture of Stenotrophomonas maltophilia (VUN 10,010), was found to also contain another bacterial species (Mycobacterium sp. strain 1B), capable of degrading a similar range of PAH substrates. Analysis of its 16S rRNA gene sequence and growth characteristics revealed the strain to be a fast-growing Mycobacterium sp., closely related to other previously isolated PAH and xenobiotic-degrading mycobacterial strains. Comparison of the PAH-degrading characteristics of Mycobacterium sp. strain 1B with those of S. maltophilia indicated some similarities (ability to degrade phenanthrene and pyrene), but some differences were also noted (S. maltophilia able to degrade fluorene, but not fluoranthene, whereas Mycobacterium sp. strain 1B can degrade fluoranthene, but not fluorene). Unlike the S. maltophilia culture, there was no evidence of benzo[a]pyrene degradation by Mycobacterium sp. strain 1B, even in the presence of other PAHs (ie pyrene) as co-metabolic substrates. Growth of Mycobacterium sp. strain 1B on other organic carbon sources was also limited compared with the S. maltophilia culture. Conclusions:, This study isolated a Mycobacterium strain from a bacterial culture capable of benzo[a]pyrene cometabolism. The Mycobacterium strain displays different PAH-degrading characteristics to those described previously for the PAH-degrading bacterial culture. It is unclear what role the two bacterial strains play in benzo[a]pyrene cometabolism, as the Mycobacterium strain does not appear to have endogenous benzo[a]pyrene degrading ability. Significance and Impact of the Study:, This study describes the isolation and characterization of a novel PAH-degrading Mycobacterium strain from a PAH-degrading culture. Further studies utilizing this strain alone, and in combination with other members of the consortium, will provide insight into the diverse roles different bacteria may play in PAH degradation in mixed cultures and in the environment. [source] Integrated polymerase chain reaction-based procedures for the detection and identification of species and subspecies of the Gram-positive bacterial genus LactococcusJOURNAL OF APPLIED MICROBIOLOGY, Issue 2 2002Z.Y. Pu Aims:,Five species of the Gram-positive bacterial genus Lactococcus (Lactococcus lactis, L. garvieae, L. plantarum, L. piscium and L. raffinolactis) are currently recognized. The aim of this work was to develop a simple approach for the identification of these species, as well as to differentiate the industrially important dairy subspecies L. lactis subsp. lactis and L. lactis subsp. cremoris. Methods and Results:,Methods were devised based on specific polymerase chain reaction (PCR) amplifications that exploit differences in the sequences of the 16S ribosomal RNA genes of each species, followed by restriction enzyme cleavage of the PCR products. The techniques developed were used to characterize industrial cheese starter strains of L. lactis and the results were compared with biochemical phenotype and DNA sequence data. Conclusions:,The PCR primers designed can be used simultaneously, providing a simple scheme for screening unknown isolates. Strains of L. lactis show heterogeneity in the 16S ribosomal RNA gene sequence. Significance and Impact of the Study:,This work provides an integrated set of methods for differentiation and identification of lactococcal species associated with agricultural, veterinary, medical and processed food industries. [source] Lancefield group C Streptococcus dysgalactiae infection responsible for fish mortalities in JapanJOURNAL OF FISH DISEASES, Issue 12 2004R Nomoto Abstract A Lancefield serological group C Streptococcus sp. was isolated from cultured amberjack, Seriola dumerili Risso, and yellowtail, Seriola quinqueradiata Temminck and Schlegel, immunized with Lactococcus garvieae commercial vaccines in Japan. The isolated bacteria were Gram-positive cocci, auto-aggregating in saline, morphologically long chains in growth medium, catalase negative and , -haemolytic on blood agar. An almost complete gene sequence of the 16S rDNA of two isolates was determined and compared with that of bacterial strains in the database. The isolates were identified as Streptococcus dysgalactiae based on the results of the 16S rDNA sequence, the bacteriological properties and the Lancefield serological grouping. Oligonucleotide primers specifically designed for the 16S,23S rDNA intergenic spacer region of S. dysgalactiae amplified a gene from all the fish isolates, as well as the type strains , -haemolytic S. dysgalactiae subsp. dysgalactiae ATCC430738 and , -haemolytic S. dysgalactiae subsp. equisimilis ATCC35666, but not those of S. equi ATCC33398, Lactococcus garvieae ATCC43921 and L. garvieae KG9408. The severe necrotic lesions of the caudal peduncle seen in experimentally infected fish were similar to those seen in naturally infected fish. [source] Molecular characterization of VP4, VP6, VP7, NSP4, and NSP5/6 genes identifies an unusual G3P[10] human rotavirus strainJOURNAL OF MEDICAL VIROLOGY, Issue 1 2009Pattara Khamrin Abstract An unusual strain of human rotavirus G3P[10] (CMH079/05) was detected in a stool sample of a 2-year-old child admitted to the hospital with severe diarrhea in Chiang Mai, Thailand. Analysis of the VP7 gene sequence revealed highest identities with unusual human rotavirus G3 strain CMH222 at 98.7% on the nucleotide and 99.6% on the amino acid levels. Phylogenetic analysis of the VP7 sequence confirmed that the CMH079/05 strain formed a cluster with G3 rotavirus reference strains and showed the closest lineage with the CMH222 strain. Analysis of partial VP4 gene of CMH079/05 revealed highest degree of sequence identities with P[10] rotavirus prototype strain 69M at nucleotide and amino acid levels of 92.9% and 94.6%, respectively. Phylogenetic analysis of the VP4 sequence revealed that CMH079/05 and 69M clustered closely together in a monophyletic branch separated from other rotavirus genotypes. To our knowledge, this is a novel G,P combination of G3 and P[10] genotypes. In addition, analyses of VP6, NSP4, and NSP5/6 genes revealed these uncommon genetic characteristics: (i) the VP6 gene differed from the four other known subgroups; (ii) the NSP4 gene was identified as NSP4 genetic group C, an uncommon group in humans; and (iii) the NSP5/6 gene was most closely related with T152, a G12P[9] rotavirus previously isolated in Thailand. The finding of uncommon G3P[10] rotavirus in this pediatric patient provided additional evidence of the genetic diversity of human group A rotaviruses in Chiang Mai, Thailand. J. Med. Virol. 81:176,182, 2009. © 2008 Wiley-Liss, Inc. [source] Isolation and functional analysis of five HPVE6 variants with respect to p53 degradationJOURNAL OF MEDICAL VIROLOGY, Issue 3 2008Thomas Hiller Abstract Persistent infection with high risk human papillomavirus is a necessary risk factor in the etiology of invasive cervical carcinoma. With regard to molecular details, the best studied types are HPV16 and HPV18 which are found in 70% of cervical cancer worldwide, however factors associated with the progression of individual cervical intraepithelial neoplasias into cancer are still poorly understood. Intratype amino acid variations in the immortalizing and transforming early proteins E6 and E7 were described to be associated with progressive disease and linked to increased viral persistence or progression. One of the key actions of high risk HPVE6 proteins is the inhibition of the function of p53, a tumor suppressor protein, by enhancing its degradation through the ubiquitin pathway. In this study, variants of five HPV type E6 proteins (HPV35, 53, 56, 66, and 70) isolated from patient materials are described and functional analysis of them were done with respect to p53 degradation. Interestingly the E6 protein of HPV type 53, which has no consistent risk classification in the literature showed the highest variability in our study. The analysis of all variants revealed no differences with regard to the degradation ability for p53 compared to the prototype E6 proteins, suggesting that the variants tested revealed no altered functions related to the carcinogenicity of the respective HPV types. It therefore seems more likely that variations in the E6 gene sequence may allow evasion from the hosts immune system, supporting increased viral persistence. J. Med. Virol. 80:478,483, 2008. © 2008 Wiley-Liss, Inc. [source] Rapid diversification of measles virus genotypes circulating in Morocco during 2004,2005 epidemics,JOURNAL OF MEDICAL VIROLOGY, Issue 11 2006Amal Alla Abstract Measles virus strains circulating in six different regions in Morocco during 2004,2005 were analysed. They were genotyped using two different methods: the recently developed method based on real-time PCR amplification and melting curve analyses, and the conventional method based on nucleic acid sequencing and phylogenetic analysis of 456 nucleotides of the 3,-region of the nucleoprotein (N) gene sequence. Five genotypes (A, B3.2, C2, D7 and D8) were shown to be circulating during this period. Previous studies on measles virus genotypes in Morocco (1998,2003) showed that only the genotype C2 was present and was considered to be endemic. Sequence comparison of the 2004,2005 viruses with other measles strains suggests that measles strains belonging to genotype B3.2 were probably imported from West Africa, whereas those belonging to genotypes D7 and D8 were imported from Europe. These studies which identify the route of importation of measles are important for developing strategies for measles elimination in Morocco. J. Med. Virol. 78:1465,1472, 2006. © 2006 Wiley-Liss, Inc. [source] Co-circulation of two genotypes of measles virus and mutual change of the prevailing genotypes every few years in Osaka, JapanJOURNAL OF MEDICAL VIROLOGY, Issue 2 2003Hideyuki Kubo Abstract Genotypes of 44 wild-type measles virus (MV) strains isolated in Osaka, Japan, during 1997,2001, were determined based on phylogenetic analyses of a 456-nt 3, terminal nucleoprotein gene sequence with the reference MV strains designated by the World Health Organization. The wild-type MV strains were classified into two genotypes, D3 and D5, recognized as indigenous in Japan. Six of 12 strains isolated in 1997 were classified into genotype D3 and the other 6 into D5. Eleven of 13 strains were D3, and 2 were D5 in 1998. There were no measles epidemics, and no strains were isolated in 1999. Nine of 10 strains were genotype D5, and only one was D3 in 2000, and 9 of 9 were D5 in 2001. These results indicate that the wild-type MV strains classified into genotypes D3 and D5 co-circulated without the complete change of the MV genotype in Osaka, except in 2001. Furthermore, the prevailing genotype was different between 1998 and 2000,2001. Together with a previous report about MV genotype in this area during 1993,1995, these results suggest that the mutual change of the prevailing wild-type MV genotypes between D3 and D5 occurs every few years in Osaka, Japan. J. Med. Virol. 69:273,278, 2003. © 2003 Wiley-Liss, Inc. [source] In Vivo Gene Transfer Studies on the Regulation and Function of the Vasopressin and Oxytocin GenesJOURNAL OF NEUROENDOCRINOLOGY, Issue 2 2003D. Murphy Abstract Novel genes can be introduced into the germline of rats and mice by microinjecting fertilized one-cell eggs with fragments of cloned DNA. A gene sequence can thus be studied within the physiological integrity of the resulting transgenic animals, without any prior knowledge of its regulation and function. These technologies have been used to elucidate the mechanisms by which the expression of the two genes in the locus that codes for the neuropeptides vasopressin and oxytocin is confined to, and regulated physiologically within, specific groups of neurones in the hypothalamus. A number of groups have described transgenes, derived from racine, murine and bovine sources, in both rat and mouse hosts, that mimic the appropriate expression of the endogenous vasopressin and genes in magnocellular neurones (MCNs) of the supraoptic and paraventricular nuclei. However, despite considerable effort, a full description of the cis -acting sequences mediating the regulation of the vasopressin-oxytocin locus remains elusive. Two general conclusions have nonetheless been reached. First, that the proximal promoters of both genes are unable to confer any cell-specific regulatory controls. Second, that sequences downstream of the promoter, within the structural gene and/or the intergenic region that separates the two genes, are crucial for appropriate expression. Despite these limitations, sufficient knowledge has been garnered to specifically direct the expression of reporter genes to vasopressin and oxytocin MCNs. Further, it has been shown that reporter proteins can be directed to the regulated secretory pathway, from where they are subject to appropriate physiological release. The use of MCN expression vectors will thus enable the study of the physiology of these neurones through the targeted expression of biologically active molecules. However, the germline transgenic approach has a number of limitations involving the interpretation of phenotypes, as well as the large cost, labour and time demands. High-throughput somatic gene transfer techniques, principally involving the stereotaxic injection of hypothalamic neuronal groups with replication-deficient adenoviral vectors, are now being developed that obviate these difficulties, and which enable the robust, long-lasting expression of biologically active proteins in vasopressin and oxytocin MCNs. [source] HETEROGENEITY OF THE CYANOBACTERIAL GENUS SYNECHOCYSTIS AND DESCRIPTION OF A NEW GENUS, GEMINOCYSTIS,JOURNAL OF PHYCOLOGY, Issue 4 2009Jana Korelusová The study and revision of the unicellular cyanobacterial genus Synechocystis was based on the type species S. aquatilis Sauv. and strain PCC 6803, a reference strain for this species. Uniformity in rRNA gene sequence, morphology, and ultrastructure was observed in all available Synechocystis strains, with the exception of the strain PCC 6308, which has been considered by some to be a model strain for Synechocystis. This strain differs substantially from the typical Synechocystis cluster according to both molecular (<90% of similarity, differences in 16S,23S rRNA internal transcribed spacer [ITS] secondary structure) and phenotypic criteria (different ultrastructure of cells). This strain is herein classified into the new genus Geminocystis gen. nov., as a sister taxon to the genus Cyanobacterium. Geminocystis differs from Cyanobacterium by genetic position (<94.4% of similarity) and more importantly by its different type of cell division. Because strain PCC 6308 was designated as a reference strain of the Synechocystis cluster 1 in Bergey's Manual, the members of this genetic cluster have to be revised and reclassified into Geminocystis gen. nov. Only the members of the Synechocystis cluster 2 allied with PCC 6803 correspond both genetically and phenotypically to the type species of the genus Synechocystis (S. aquatilis). [source] Sensitive and Specific Digoxigenin-labelled RNA Probes for Routine Detection of Citrus tristeza virus by Dot-blot HybridizationJOURNAL OF PHYTOPATHOLOGY, Issue 6 2006L. Barbarossa Abstract A non-radioactive dot-blot hybridization assay for the successful detection of Citrus tristeza virus (CTV) RNA in total nucleic acid extracts of infected citrus was developed. Two digoxigenin (DIG)-labelled minus-sense riboprobes, complementary to the coat protein gene sequence of a Chinese and an Apulian CTV isolate were synthesized. Several citrus tissues were evaluated as optimal virus source and leaf petioles were found appropriate material for reliable detection. The hybridization assay showed a detection limit corresponding to 0.2 mg of fresh infected tissue. The riboprobes allowed CTV detection in isolates from different geographical areas, grown in the screenhouse or in the field, resulting in similar hybridization patterns. The infected trees were tested during different seasons with positive results, although from July to August most of the samples gave a weaker hybridization signal, compared to other seasons. The high sensitivity and reliability of the molecular hybridization assay described make it a good alternative to serological methods for CTV detection. [source] Characterization of the Coat Protein Gene of Cymbidium mosaic virus Isolates from IndiaJOURNAL OF PHYTOPATHOLOGY, Issue 5 2006A. R. Sherpa Abstract The variability in the coat protein (CP) gene sequence of Cymbidium mosaic virus (CymMV) that naturally infects orchids worldwide was investigated. Samples were collected from different regions of India, and the gene encoding the CP of nine isolates was specifically amplified by reverse-transcription polymerase chain reaction. The amplified product obtained was cloned, sequenced and multiple sequence alignment of deduced amino acid (aa) sequences revealed considerable homology to CymMV isolates from other countries. The nucleotide sequences and the amino acid sequences were found to be 85,100% identical and 65,100% respectively. Such high sequence conservation suggests that the CymMV CP gene is highly conserved and is a suitable candidate for the development of diagnostic procedures and to provide transgenic resistance to orchids cultivated in different geographical locations. Although recombination is not common among CymMV isolates, one isolate from Cymbidium was found to be a recombinant between a Korean and a Thai isolate of the virus. IHBT communication no: 0451. [source] | ||||||||||||||||||||||||||||||||||||||||||||||