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Genetic Transformation (genetic + transformation)
Kinds of Genetic Transformation Selected AbstractsBiological systems of the host cell involved in Agrobacterium infectionCELLULAR MICROBIOLOGY, Issue 1 2007Vitaly Citovsky Summary Genetic transformation of plants by Agrobacterium, which in nature causes neoplastic growths, represents the only known case of trans -kingdom DNA transfer. Furthermore, under laboratory conditions, Agrobacterium can also transform a wide range of other eukaryotic species, from fungi to sea urchins to human cells. How can the Agrobacterium virulence machinery function in such a variety of evolutionarily distant and diverse species? The answer to this question lies in the ability of Agrobacterium to hijack fundamental cellular processes which are shared by most eukaryotic organisms. Our knowledge of these host cellular functions is critical for understanding the molecular mechanisms that underlie genetic transformation of eukaryotic cells. This review outlines the bacterial virulence machinery and provides a detailed discussion of seven major biological systems of the host cell,cell surface receptor arrays, cellular motors, nuclear import, chromatin targeting, targeted proteolysis, DNA repair, and plant immunity , thought to participate in the Agrobacterium -mediated genetic transformation. [source] Natural transformation of Vibrio fischeri requires tfoX and tfoYENVIRONMENTAL MICROBIOLOGY, Issue 8 2010Amber Pollack-Berti Summary Recent evidence has indicated that natural genetic transformation occurs in Vibrio cholerae, and that it requires both induction by chitin oligosaccharides, like chitohexaose, and expression of a putative regulatory gene designated tfoX. Using sequence and phylogenetic analyses we have found two tfoX paralogues in all sequenced genomes of the genus Vibrio. Like V. cholerae, when grown in chitohexaose, cells of V. fischeri are able to take up and incorporate exogenous DNA. Chitohexaose-independent transformation by V. fischeri was observed when tfoX was present in multicopy. The second tfoX paralogue, designated tfoY, is also required for efficient transformation in V. fischeri, but is not functionally identical to tfoX. Natural transformation of V. fischeri facilitates rapid transfer of mutations across strains, and provides a highly useful tool for experimental genetic manipulation in this species. The presence of chitin-induced competence in several vibrios highlights the potential for a conserved mechanism of genetic exchange across this family of environmentally important marine bacteria. [source] Regulation of natural genetic transformation and acquisition of transforming DNA in Streptococcus pneumoniaeFEMS MICROBIOLOGY REVIEWS, Issue 3 2009Ola Johnsborg Abstract The ability of pneumococci to take up naked DNA from the environment and permanently incorporate the DNA into their genome by recombination has been exploited as a valuable research tool for 80 years. From being viewed as a marginal phenomenon, it has become increasingly clear that horizontal gene transfer by natural transformation is a powerful mechanism for generating genetic diversity, and that it has the potential to cause severe problems for future treatment of pneumococcal disease. This process constitutes a highly efficient mechanism for spreading ,-lactam resistance determinants between streptococcal strains and species, and also threatens to undermine the effect of pneumococcal vaccines. Fortunately, great progress has been made during recent decades to elucidate the mechanism behind natural transformation at a molecular level. Increased insight into these matters will be important for future development of therapeutic strategies and countermeasures aimed at reducing the spread of hazardous traits. In this review, we focus on recent developments in our understanding of competence regulation, DNA acquisition and the role of natural transformation in the dissemination of virulence and ,-lactam resistance determinants. [source] Somatic embryogenesis and multiple-shoot formation from seed-derived shoot apical meristems of rhodesgrass (Chloris gayana Kunth)GRASSLAND SCIENCE, Issue 3 2007Takahiro Gondo Abstract High-frequencies of somatic embryogenesis and multiple-shoot formation were achieved with seed-derived shoot apical meristems of rhodesgrass (Chloris gayana Kunth). Shoot apices as initial explants were isolated from aseptically germinated seedlings, and cultured in vitro. Embryogenic calli and the multiple shoots were induced and maintained on MS basal medium with various combinations of 2,4-dichlorophnoxyacetic acid (2,4-D) and 6-benzylaminopurine (BAP), the two most effective treatments being 2.0 mg L,1 2,4-D and 0.1 mg L,1 2,4-D + 2.0 mg L,1 BAP. These embryogenic calli and multiple-shoot formations could be used as alternative regenerable target tissues for genetic transformation using particle bombardment of rhodesgrass. [source] Ultrathin polymeric interpenetration network with separation performance approaching ceramic membranes for biofuelAICHE JOURNAL, Issue 1 2009Lan Ying Jiang Abstract Biofuel has emerged as one of the most strategically important sustainable fuel sources. The success of biofuel development is not only dependent on the advances in genetic transformation of biomass into biofuel, but also on the breakthroughs in separation of biofuel from biomass. The "separation" alone currently accounts for 60,80% of the biofuel production cost. Ceramic membranes made of sophisticated processes have shown separation performance far superior to polymeric membranes, but suffers fragility and high fabrication cost. We report the discovery of novel molecular engineering and membrane fabrication that can synergistically produce polymeric membranes exhibiting separation performance approaching ceramic membranes. The newly discovered Polysulfone/Matrimid composite membranes are fabricated by dual-layer coextrusion technology in just one step through phase inversion. An ultrathin dense-selective layer made of an interpenetration network of the two materials with a targeted and stable interstitial space is formed at the interface of two layers for biofuel separation. The combined molecular engineering and membrane fabrication approach may revolutionize future membrane research and development for purification and separation in energy, environment, and pharmaceuticals. © 2008 American Institute of Chemical Engineers AIChE J, 2009 [source] MOLECULAR GENETIC MANIPULATION OF THE DIATOM THALASSIOSIRA PSEUDONANA (BACILLARIOPHYCEAE),JOURNAL OF PHYCOLOGY, Issue 5 2006Nicole Poulsen Here, we describe the first system for genetic transformation of Thalassiosira pseudonana (Hustedt) Hasle et Heimdal, the only diatom for which a complete genome sequence is presently available. This method is based on microparticle bombardment followed by selection of transformants using the antibiotic nourseothricin. It exhibits the highest transformation efficiency compared with transformation systems for other diatom species. To achieve the high transformation efficiency, it is important to allow recovery of the bombarded T. pseudonana cells in non-selective suspension culture before spreading on nourseothricin containing agar plates. It is demonstrated that T. pseudonana is readily susceptible to co-transformation allowing for the simultaneous introduction of a non-selective gene together with the selection marker gene. Both introduced genes are stably inherited even in the absence of the antibiotic selection pressure. We have developed two T. pseudonana -specific expression vectors that can drive constitutive expression (vector pTpfcp) and inducible expression (vector pTpNR) of introduced genes. In combination with the available genome data the T. pseudonana transformation system is expected to provide a powerful tool for functional genomics in diatoms. [source] A food-grade site-directed mutagenesis system for Streptococcus thermophilus LMG 18311LETTERS IN APPLIED MICROBIOLOGY, Issue 3 2010T. Blomqvist Abstract Aims:, To develop a general method for site-directed mutagenesis in the dairy starter strain Streptococcus thermophilus LMG 18311 which does not depend on antibiotic-resistance genes or other selection markers for the identification of transformants. Methods and Results:, In a previous study, we demonstrated that Strep. thermophilus LMG 18311 can be made competent for natural genetic transformation by overexpression of the alternative sigma factor ComX. In the present study, we wanted to investigate whether the natural transformation mechanism of Strep. thermophilus LMG 18311 is efficient enough to make it feasible to perform site-directed mutagenesis in this strain without the use of a selection marker. Competent bacteria were mixed with a DNA fragment engineered to contain a nonsense and a frameshift mutation in the middle of the target gene (lacZ) and subsequently seeded on agar plates. By performing colony-lift hybridization using a digoxigenin-labelled oligonucleotide probe, we succeeded in identifying transformants containing the sought after mutation. Conclusions:, By exploiting the natural transformability of Strep. thermophilus LMG 18311 and standard molecular methods, we have demonstrated that the genome of this bacterium can be altered at preselected sites without introduction of any foreign DNA. Significance and Impact of the Study:, A food-grade site-directed mutagenesis system has been developed for Strep. thermophilus LMG 18311 that can be used by the dairy industry to construct starter strains with novel and/or improved properties. [source] Expression of ,re,y luciferase gene in Erwinia amylovoraLUMINESCENCE: THE JOURNAL OF BIOLOGICAL AND CHEMICAL LUMINESCENCE, Issue 4 2003Giovanna Gentilomi Abstract In this study we describe an ef,cient stable genetic transformation of the phytopathogenic bacterium Erwinia amylovora using a recombinant expression vector encoding the ,re,y luciferase gene of Photinus pyralis, which is further controlled by IPTG-inducible promoter. Stably transformed E. amylovora cells maintain the same infectivity as the wild-type strain and, after induction with IPTG, produce luciferase. Luminescence produced by the action of luciferase on an exogenous substrate was easily detectable by a simple and rapid bioluminescent assay (BL). The transformed E. amylovora strain maintains the same high emission level, even after passage in pears, until about 15 days post-infection. Our ,ndings therefore show that the luciferase assay can be conveniently used to follow the bacterial movement in plant tissue and its dissemination in controlled environments. [source] Enabling technologies to improve area-wide integrated pest management programmes for the control of screwwormsMEDICAL AND VETERINARY ENTOMOLOGY, Issue 2009A. S. ROBINSON Abstract The economic devastation caused in the past by the New World screwworm fly Cochliomyia hominivorax (Coquerel) (Diptera: Calliphoridae) to the livestock industry in the U.S.A., Mexico and the rest of Central America was staggering. The eradication of this major livestock pest from North and Central America using the sterile insect technique (SIT) as part of an area-wide integrated pest management (AW-IPM) programme was a phenomenal technical and managerial accomplishment with enormous economic implications. The area is maintained screwworm-free by the weekly release of 40 million sterile flies in the Darien Gap in Panama, which prevents migration from screwworm-infested areas in Columbia. However, the species is still a major pest in many areas of the Caribbean and South America and there is considerable interest in extending the eradication programme to these countries. Understanding New World screwworm fly populations in the Caribbean and South America, which represent a continuous threat to the screwworm-free areas of Central America and the U.S.A., is a prerequisite to any future eradication campaigns. The Old World screwworm fly Chrysomya bezziana Villeneuve (Diptera: Calliphoridae) has a very wide distribution ranging from Southern Africa to Papua New Guinea and, although its economic importance is assumed to be less than that of its New World counterpart, it is a serious pest in extensive livestock production and a constant threat to pest-free areas such as Australia. In the 1980s repeated introductions and an expansion of Old World screwworm populations were reported in the Middle East; in the 1990s it invaded Iraq and since late 2007 it has been reported in Yemen, where a severe outbreak of myiasis occurred in 2008. Small-scale field trials have shown the potential of integrating the SIT in the control of this pest and various international organizations are considering using the release of sterile insects as part of an AW-IPM approach on a much wider scale. Wohlfahrtia magnifica (Schiner) (Diptera: Sarcophagidae) is a screwworm of temperate regions, which, although of limited agricultural importance, has invaded several new locations in the past few years. This special issue reports on the results of a 6-year project funded by the Joint Food and Agriculture Organization of the United Nations/International Atomic Energy Agency (FAO/IAEA) Programme of Nuclear Techniques in Food and Agriculture entitled ,Enabling Technologies for the Expansion of the SIT for Old and New World Screwworm'. A major goal of the project was to better understand population genetic variation in screwworms as an aid to the identification of isolated populations. The project also addressed issues related to genetic sexing, cuticular hydrocarbons, population dynamics, genetic transformation and chromosome analysis. [source] MicroReview: Competence-induced fratricide in streptococciMOLECULAR MICROBIOLOGY, Issue 6 2007Jean-Pierre Claverys Summary Competence for natural genetic transformation in Streptococcus pneumoniae is controlled by the extracellular concentration of the competence-stimulating peptide (CSP), an exported peptide pheromone. Upon entering the competent state, pneumococci start transcribing a number of CSP-responsive genes, termed the early and late competence (com) genes. Some of the proteins encoded by these com genes are absolutely required for DNA uptake and transformation, but most of them are dispensable. This finding indicates that the majority of CSP-regulated proteins in S. pneumoniae is involved in processes unrelated to natural genetic transformation. Recently, however, it became clear that the biological role of a few of the dispensable proteins might be linked to the transformation process. Although these proteins are not needed for transformation per se, they constitute a killing mechanism that could be used by competent cells to acquire DNA from non-competent pneumococci. This mechanism, termed fratricide, has so far only been described for pneumococci. In this manuscript, we review evidence that suggests the conservation of fratricide as well as the independent evolution of its genetic control and of its effectors in several species of the genus Streptococcus, and discuss its possible biological significance in relation to natural transformation. [source] Ethylene production in plants during transformation suppresses vir gene expression in Agrobacterium tumefaciensNEW PHYTOLOGIST, Issue 3 2008Satoko Nonaka Summary ,,Ethylene evolution from plants inhibits Agrobacterium -mediated genetic transformation, but the mechanism is little understood. In this study, the possible role of ethylene in Agrobacterium -mediated genetic transformation was clarified. ,,It was tested whether or not plant ethylene sensitivity affected genetic transformation; the sensitivity might regulate bacterial growth during co-cultivation and vir gene expression in Agrobacterium tumefaciens. For these experiments, melon (Cucumis melo) was used, in which ethylene sensitivity was controlled by chemicals, and Arabidopsis ethylene-insensitive mutants. ,,Agrobacterium -mediated genetic transformation was inhibited in ethylene-sensing melon, whereas, in Arabidopsis ethylene-insensitive mutant, it was enhanced. However, the ethylene sensitivity did not affect bacterial growth. vir gene expression was inhibited by application of plant exudate from ethylene-sensitive plants. The inhibitory effect of the ethylene sensitivity on genetic transformation relieved the activation of vir gene expression in A. tumefaciens with vir gene inducer molecule (acetosyringone, AS) or A. tumefaciens mutant strain which has constitutive vir gene expression. ,,These results indicate that ethylene evolution from a plant inoculated with A. tumefaciens inhibited vir gene expression in A. tumefaciens through the ethylene signal transduction in the plant, and, as a result, Agrobacterium -mediated genetic transformation was inhibited. [source] Biotechnology of the Banana: A Review of Recent ProgressPLANT BIOLOGY, Issue 5 2000G. R. Rout Abstract: A number of biotechnological tools have been developed which could help breeders to evolve new plant types to meet the demand of the food industry in the next century. Available techniques for the transfer of genes could significantly shorten the breeding procedures and overcome some of the agronomic and environmental problems which would otherwise not be possible through conventional methods. In vitro protocols have been standardized to allow commercially viable propagation of desired clones of Musa. An overview of the regeneration of banana by direct and indirect organogenesis, and somatic embryogenesis is presented in this article. In addition, the use of several other biotechnological techniques to enrich the genome of banana, such as selection of somaclonal variants, screening for various useful characteristics, cryopreservation, genetic transformation and molecular genetics are reviewed. In conclusion, the improvement of banana through modern biotechnology should help ensure food security by stabilizing production levels in sustainable cropping systems geared towards meeting domestic and export market demands. [source] Agrobacterium -mediated transformation of cereals: a promising approach crossing barriersPLANT BIOTECHNOLOGY JOURNAL, Issue 6 2006Ashok Kumar Shrawat Summary Cereal crops have been the primary targets for improvement by genetic transformation because of their worldwide importance for human consumption. For a long time, many of these important cereals were difficult to genetically engineer, mainly as a result of their inherent limitations associated with the resistance to Agrobacterium infection and their recalcitrance to in vitro regeneration. The delivery of foreign genes to rice plants via Agrobacterium tumefaciens has now become a routine technique. However, there are still serious handicaps with Agrobacterium -mediated transformation of other major cereals. In this paper, we review the pioneering efforts, existing problems and future prospects of Agrobacterium -mediated genetic transformation of major cereal crops, such as rice, maize, wheat, barley, sorghum and sugarcane. [source] Increased cysteine availability is essential for cadmium tolerance and accumulation in Arabidopsis thalianaPLANT BIOTECHNOLOGY JOURNAL, Issue 6 2004José R. Domínguez-Solís Summary Employing genetic transformation using an Atcys-3A cDNA construct expressing the cytosolic O -acetylserine(thiol)lyase (OASTL), we obtained two Arabidopsis lines with different capabilities for supplying cysteine under metal stress conditions. Lines 1-2 and 10-10, grown under standard conditions, showed similar levels of cysteine and glutathione (GSH) to those of the wild-type. However, in the presence of cadmium, line 10-10 showed significantly higher levels. The increased thiol content allowed line 10-10 to survive under severe heavy metal stress conditions (up to 400 µm of cadmium in the growth medium), and resulted in an accumulation of cadmium in the leaves to a level similar to that of metal hyperaccumulator plants. Investigation of the epidermal leaf surface clearly showed that most of the cadmium had accumulated in the trichomes. Furthermore, line 10-10 was able to accumulate more cadmium in its trichomes than the wild-type, whereas line 1-2 showed a reduced capacity for cadmium accumulation. Our results suggest that an increased rate of cysteine biosynthesis is responsible for the enhanced cadmium tolerance and accumulation in trichome leaves. Thus, molecular engineering of the cysteine biosynthesis pathway, together with modification of the number of leaf trichomes, may have considerable potential in increasing heavy metal accumulation for phytoremediation purposes. [source] Plant regeneration through callus initiation from mature embryo of TriticumPLANT BREEDING, Issue 1 2007R. M. Bi Abstract The behaviour of diverse Triticum genotypes in the tissue culture response of mature embryo callus was compared, and factors affecting tissue culture response were studied in this paper. Significant differences were detected in callus induction, embryogenic callus differentiation, plantlet regeneration and culture efficiency when mature embryos of 31 plants of different Triticum species were compared. These were the main wheat cultivars of the Chinese northern winter-type wheat region and breeding lines (Triticum aestivum L.), durum wheat (Triticum durum Desf.), cultivable emmer wheat (Triticumdicoccum Schuble) and the common wheat progenitors Triticum dicoccoides and Triticum aegilopides. The genotype dependency was particularly high in tissue culture of mature embryos of these Triticum genotypes. The efficiency of induction, differentiation and regeneration of mature embryos callus was high in genotypes selected out. Mature embryo-derived callus of HB341, TS021, SN2618, T. dicoccum, HB188, and T9817 showed better tissue culture response than the other genotypes. Plantlets can be regenerated from mature embryo-derived callus of 31 genotypes, saving on growth facility resources and time required for the collection of other explants, and providing a solid basis for the genetic transformation and molecular plant breeding of Triticum plants. [source] Insect-symbiont systems: From complex relationships to biotechnological applicationsBIOTECHNOLOGY JOURNAL, Issue 12 2009Sandra Chaves Abstract Microbial symbiosis is a ubiquitous aspect of life and was a major element in the ability of insects to explore several adverse environments. To date, the study of symbiosis in insects has been impaired by the unculturability of most symbionts. However, some molecular methods represent powerful tools to help understand insect-microorganism associations and to disclose new symbiont-host systems. Beyond playing an essential role in nutrition and development of the insects, symbionts can produce bioactive compounds that protect the host against adverse environmental conditions, predators and/or direct competitors. Since the search for natural bioactive products and new enzymes is a developing area, understanding the diversity and nature of symbiont-host relationships paves the way for the exploitation of new resources in biotechnology. Furthermore, genetic transformation of the symbionts with genes that code for compounds that are toxic for pathogenic and phytopathogenic agents is also a promising area of application of the insect-symbiont relationships. The search for new bioactive compounds, the use of symbionts for pest and disease control and the molecular strategies applied for these purposes are issues of particular interest for innovative biotechnological applications and are addressed in the present review. [source] Evidence that direct DNA uptake through cut shoots leads to genetic transformation of Solanum aviculare ForstCELL BIOCHEMISTRY AND FUNCTION, Issue 1 2003P. B. Gahan Abstract The reporter genes GUS, NPTII and BAR, either separately or in combination, have been exploited to determine if DNA which can directly enter plants, circulate within the plant and enter nuclei, can also integrate into the genome in a manner which will permit gene expression. Feeding of either seed-derived or adventitious cut shoots of Solanum aviculare with the GUS gene followed by rooting of the shoots and growing on, resulted in all tissues of the plant showing GUS activity as detected cytochemically. Southern blot analysis of plants derived from the adventitious shoots confirmed the presence of the reporter gene in roots. Reporter gene expression was observed also in the F1 generation. If GUS and NPTII or GUS, NPTII and BAR were fed together, then in each case it was possible to have both expression and Southern blot confirmation of each of the genes. There was a relatively high rate of transformation of approximately 5% of the fed stems across all experiments conducted during the present study. Copyright © 2002 John Wiley & Sons, Ltd. [source] Biological systems of the host cell involved in Agrobacterium infectionCELLULAR MICROBIOLOGY, Issue 1 2007Vitaly Citovsky Summary Genetic transformation of plants by Agrobacterium, which in nature causes neoplastic growths, represents the only known case of trans -kingdom DNA transfer. Furthermore, under laboratory conditions, Agrobacterium can also transform a wide range of other eukaryotic species, from fungi to sea urchins to human cells. How can the Agrobacterium virulence machinery function in such a variety of evolutionarily distant and diverse species? The answer to this question lies in the ability of Agrobacterium to hijack fundamental cellular processes which are shared by most eukaryotic organisms. Our knowledge of these host cellular functions is critical for understanding the molecular mechanisms that underlie genetic transformation of eukaryotic cells. This review outlines the bacterial virulence machinery and provides a detailed discussion of seven major biological systems of the host cell,cell surface receptor arrays, cellular motors, nuclear import, chromatin targeting, targeted proteolysis, DNA repair, and plant immunity , thought to participate in the Agrobacterium -mediated genetic transformation. [source] Molecular interactions between Plasmodium and its insect vectorsCELLULAR MICROBIOLOGY, Issue 11 2002R. E. Sinden Summary Our understanding of the intricate interactions between the malarial parasite and the mosquito vector is complicated both by the number and diversity of parasite and vector species, and by the experimental inaccessibility of phenomena under investigation. Steady developments in techniques to study the parasite in the mosquito have recently been augmented by methods to culture in their entirety the sporogonic stages of some parasite species. These, together with the new saturation technologies, and genetic transformation of both parasite and vector will permit penetrating studies into an exciting and largely unknown area of parasite,host interactions, an understanding of which must result in the development of new intervention strategies. This microreview highlights key areas of current basic molecular interest, and identifies numerous lacunae in our knowledge that must be filled if we are to make rational decisions for future control strategies. It will conclude by trying to explain why in the opinion of this reviewer understanding malaria,mosquito interactions may be critical to our future attempts to limit a disease of growing global importance. [source] | ||||||||||||||||||||||||||||