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Bp Open Reading Frame (bp + open_reading_frame)
Selected AbstractsMolecular cloning of CYP1A gene and its expression by benzo(a)pyrene from goldfish (Carassius auratus)ENVIRONMENTAL TOXICOLOGY, Issue 3 2009Seung-Min Oh Abstract We cloned and sequenced the cytochrome P450 1A (CYP1A) gene from goldfish (Carassius auratus). It has a 1581 bp open reading frame that encodes a 526 amino acid protein with a theoretical molecular weight of 59.02 kDa. The CYP1A amino acid sequence clusters in a monophyletic group with other fish CYP1As, and more closely related to zebrafish CYP1A (91% identity) than to other fish CYP1As. Exposure to benzo(a)pyrene (BaP) by intraperitoneal injection increased biliary BaP metabolites and liver CYP1A gene expression. BaP exposure also increased CYP1A gene expression in extrahepatic organs, including intestine, and gill, which are sensitive to aqueous and dietary exposure to Arylhydrocarbon receptor (AhR) agonists. Therefore, goldfish CYP1A identified in this study offers basic information for further research related to biomarker use of CYP1A of goldfish. © 2008 Wiley Periodicals, Inc. Environ Toxicol, 2009. [source] Cloning, sequencing, and characterization of CYP1A1 cDNA from leaping mullet (Liza Saliens) liver and implications for the potential functions of its conserved amino acidsJOURNAL OF BIOCHEMICAL AND MOLECULAR TOXICOLOGY, Issue 5 2001Alaattin Sen Abstract A 2,037 bp CYP1A1 cDNA (GenBank AF072899) was cloned through screening of a ,ZipLox cDNA library constructed from the liver of a leaping mullet (Liza saliens) fish captured from Izmir Bay on the Aegean coast of Turkey using rainbow trout CYP1A1 cDNA as a probe. This clone has a 130 bp 5'-flanking region, a 1,563 bp open reading frame (ORF) encoding a 521-amino acid protein (58,972 Da), and a 344 bp 3'-untranslated region without a poly (A) tail. Alignment of the deduced amino acids of CYP1A1 cDNAs showed 58% and 69,96% identities with human and 12 other fish species, respectively. Southern blot analysis suggested that this CYP1A1 cDNA was from a single-copy gene. Based on the comparison with CYP1A1 genes reported for fish and mammals, the leaping mullet CYP1A1 gene is probably split into 7 exons. The intron insertion sites were predicted. Alignment of the CYP1A1 cDNA encoded amino acids from 13 fish and 7 mammalian species disclosed differences in highly conserved amino acids between aquatic and land vertebrates. The possible associated secondary structure; conserved motifs and substrate-binding sites were discussed. The phylogenetic relationships of CYP1A1s among 13 fish species were analyzed by a distance method. © 2001 John Wiley & Sons, Inc. J Biochem Mol Toxicol 15:243,255, 2001 [source] W55a Encodes a Novel Protein Kinase That Is Involved in Multiple Stress ResponsesJOURNAL OF INTEGRATIVE PLANT BIOLOGY, Issue 1 2009Zhao-Shi Xu Abstract Protein kinases play crucial roles in response to external environment stress signals. A putative protein kinase, W55a, belonging to SNF1-related protein kinase 2 (SnRK2) subfamily, was isolated from a cDNA library of drought-treated wheat seedlings. The entire length of W55a was obtained using rapid amplification of 5, cDNA ends (5,-RACE) and reverse transcription-polymerase chain reaction(RT-PCR). It contains a 1 029 -bp open reading frame (ORF) encoding 342 amino acids. The deduced amino acid sequence of W55a had eleven conserved catalytic subdomains and one Ser/Thr protein kinase active-site that characterize Ser/Thr protein kinases. Phylogenetic analysis showed that W55a was 90.38% homologous with rice SAPK1, a member of the SnRK2 family. Using nullisomic-tetrasomic and ditelocentric lines of Chinese Spring, W55a was located on chromosome 2BS. Expression pattern analysis revealed that W55a was upregulated by drought and salt, exogenous abscisic acid, salicylic acid, ethylene and methyl jasmonate, but was not responsive to cold stress. In addition, W55a transcripts were abundant in leaves, but not in roots or stems, under environmental stresses. Transgenic Arabidopsis plants overexpressing W55a exhibited higher tolerance to drought. Based on these findings, W55a encodes a novel dehydration-responsive protein kinase that is involved in multiple stress signal transductions. [source] Molecular cloning and characterization of Bombyx mori CREB gene,ARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY (ELECTRONIC), Issue 1 2009Hongsheng Song Abstract The cAMP response element binding protein (CREB), as one of the best characterized stimulus-induced transcription factors, plays critical roles in activating transcription of target genes in response to a variety of environmental stimuli. To characterize this important molecule in the silkworm, Bombyx mori, we cloned a full-length cDNA of CREB gene from B. mori brains by using RACE-PCR. The sequence of B. mori CREB (named BmCREB1) gene contains a 88,bp 5, UTR, a 783,bp open reading frame (ORF) encoding 261 amino acids and a 348,bp 3, UTR. The deduced BmCREB amino acid sequence has 56.7% and 37.2% homology with CREB from Apis mellifera carnica and Drosophila melanogaster, respectively. The primary structure of the deduced BmCREB1 protein contains a kinase-inducible domain (KID) and a basic region/leucine zipper (bZIP) dimerization domain which exisits in all CREB family members. Genomic analysis showed there are 9 exons and 5 introns in B. mori CREB genome sequences. We identified three different isoforms of BmCREB (BmCREB1, BmCREB2 and BmCREB3) through alternative splicing in C terminal. In addition, the expression of BmCREB in different developmental stages was investigated by using quantitative real-time PCR in both diapause and non-diapause type of B. mori bivoltine race (Dazao). BmCREB transcripts showed two peaks in embryonic stage and pupal stage in both types of bivoltine race. However, consistently higher expression of BmCREB was found throughout the developmental stages in the diapause type than in the non-diapause type. These results suggest that BmCREB is involved in the processs of diapause induced by environmental factors. © 2009 Wiley Periodicals, Inc. [source] Cloning and characterization of a 70 kDa heat shock cognate gene (HSC70) from two species of ChironomusINSECT MOLECULAR BIOLOGY, Issue 1 2003N. K. Karouna-Renier Abstract In the present study we carried out the isolation and characterization of an HSC70 gene from two midges, Chironomus tentans and C. yoshimatsui. The HSC70 cDNAs are approximately 2424 (C. tentans) and 2464 bp (C. yoshimatsui) long, and contain 1950 and 1956 bp open reading frames, respectively. Analysis of genomic DNA revealed the presence of two introns in these genes. The 5, untranslated regions of the HSC70 genes are adenosine-rich, a feature found in inducible HSP70 genes. The nucleotide and amino acid sequences exhibit high identity with cytosolic HSC70s from other Dipterans. Northern hybridization indicated that HSC70 is expressed at all developmental stages, from embryo to adult, and Southern hybridization confirmed the presence of multiple HSP70 genes in Chironomus. [source] Differential mRNA expression levels and gene sequences of carboxylesterase in both deltamethrin resistant and susceptible strains of the cotton aphid, Aphis gossypiiINSECT SCIENCE, Issue 3 2008Chuan-Wang Cao Abstract Extensive use of insecticides on cotton has prompted resistance development in the cotton aphid, Aphis gossypii (Glover) in China. A deltamethrin-selected population of cotton aphids from Xinjiang Uygur Autonomous Region, China with 228.59-fold higher resistance to deltamethrin was used to examine how carboxylesterase conferred resistance to this pyrethroid insecticide. The carboxylesterase activity in the deltamethrin-resistant strain was 3.67-, 2.02- and 1.16-fold of the susceptible strain when using ,-naphthyl acetate (,-NA), ,-naphthyl acetate (,-NA) and ,-naphthyl butyrate (,-NB) as substrates, respectively. Carboxylesterase cDNA was cloned and sequenced from both deltamethrin-resistant and susceptible strains. The cDNA contained 1581 bp open reading frames (ORFs) coding a 526 amino acid protein. Only one amino acid substitution (Val87 -Ala) was observed between deltamethrin-resistant and susceptible strains but it is not genetically linked to resistance by the catalytic triad and signature motif analysis. The real-time polymerase chain reaction analysis indicated that the resistant strain had a 6.61-fold higher level of carboxylesterase mRNA than the susceptible strain. The results revealed that up-regulation of the carboxylesterase gene, not modified gene structure, may be responsible for the development of resistance in cotton aphids to deltamethrin. [source] | |||||||||||||