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Yan Li Il-Chan Kim Young Ja Kim Moon Kyoo Kim Yong-Dal Yoon Yong-Sung Lee Jae-Seong Lee 《Marine environmental research》2004,58(2-5):681
We have cloned the proto-oncogene c-fos from a self-fertilizing fish Rivulus marmoratus (Cyprinodontiformes, Rivulidae) after screening of R. marmoratus λGEM-11 genomic DNA library, and sequenced over 12 kb including all exons, introns and the promoter region. The R. marmoratus c-fos gene consisted of one noncoding exon and four exons with high similarity to those of fugu and mammals. We sequenced ≈7 kb of the R. marmoratus c-fos gene promoter region to gain a better understanding of the molecular anatomy of the immediate response of this gene upon cellular damage. In the promoter region, R. marmoratus c-fos gene has seven xenobiotic response elements (XREs) and eight metal response elements (MREs) as well as two estradiol (E2), 4 NFκB, 2 CarG, 2 prolactin (PRL) motifs and one pit1 site, while the 3′-UTR of this gene contains the estrogen response element (ERE). The seven XRE and eight MRE motifs raise the possibility of its regulation by exposure to environmental pollutants. In this paper, we discuss the gene structure of R. marmoratus c-fos gene and compare its promoter region with those of other organisms' c-fos genes. We propose its potential use in ecotoxicology. 相似文献
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Kim IC Chang SY Williams TD Ja Kim Y Yoon YD Lee YS Park EH Lee JS 《Marine environmental research》2004,58(2-5):687-691
We cloned the vitellogenin gene from the self-fertilizing fish Rivulus marmoratus, and sequenced 12,326 bp. The number of exons of R. marmoratus and rainbow trout vitellogenin genes were different, and also the splicing junctions are different throughout most of the exons and introns but the amino acid similarity of R. marmoratus vitellogenin gene to other species was rather high. In promoter region of R. marmoratus vitellogenin gene, there were several E2 binding sites and the estrogen response element (ERE). We discuss here the gene structure and expression of R. marmoratus vitellogenin gene. 相似文献
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To use two small fish Rivulus marmoratus (Cyprinodontiformes, Rivulidae) and the Japanese medaka Oryzias latipes (Belloniformes) as testing models in molecular ecotoxicology, we have cloned the cytochrome P450 1A (CYP1A) gene after screening of both genomic DNA libraries, and sequenced 11,863 and 7,243 bp including all the exons and introns with promoter regions, respectively. The Rivulus and the medaka CYP1A gene consisted of seven exons (including non-coding exons) with high homology to mammals. In the promoter region, Rivulus CYP1A gene has seven xenobiotic response elements (XREs) and two metal response elements (MREs), while the Japanese medaka CYP1A gene has six XREs and four MREs. Interestingly, medaka CYP1A gene has a number of MREs at the promoter, which may affect its response on metal exposure. We describe here the gene structure of both fish CYP1A genes. 相似文献
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Il-Chan Kim Young Ja Kim Yong-Dal Yoon Shoji Kawamura Yong-Sung Lee Jae-Seong Lee 《Marine environmental research》2004,58(2-5):125
To use two small fish Rivulus marmoratus (Cyprinodontiformes, Rivulidae) and the Japanese medaka Oryzias latipes (Belloniformes) as testing models in molecular ecotoxicology, we have cloned the cytochrome P450 1A (CYP1A) gene after screening of both genomic DNA libraries, and sequenced 11,863 and 7,243 bp including all the exons and introns with promoter regions, respectively. The Rivulus and the medaka CYP1A gene consisted of seven exons (including non-coding exons) with high homology to mammals. In the promoter region, Rivulus CYP1A gene has seven xenobiotic response elements (XREs) and two metal response elements (MREs), while the Japanese medaka CYP1A gene has six XREs and four MREs. Interestingly, medaka CYP1A gene has a number of MREs at the promoter, which may affect its response on metal exposure. We describe here the gene structure of both fish CYP1A genes. 相似文献
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Il-Chan Kim Sung Yeoul Chang Tim D. Williams Young Ja Kim Yong-Dal Yoon Yong-Sung Lee Eun-Ho Park Jae-Seong Lee 《Marine environmental research》2004,58(2-5):687
We cloned the vitellogenin gene from the self-fertilizing fish Rivulus marmoratus, and sequenced 12,326 bp. The number of exons of R. marmoratus and rainbow trout vitellogenin genes were different, and also the splicing junctions are different throughout most of the exons and introns but the amino acid similarity of R. marmoratus vitellogenin gene to other species was rather high. In promoter region of R. marmoratus vitellogenin gene, there were several E2 binding sites and the estrogen response element (ERE). We discuss here the gene structure and expression of R. marmoratus vitellogenin gene. 相似文献
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Concensus primers designed to CYP1A-conserved regions were used to amplify a 1.3 kb probe from flounder genomic DNA via polymerase chain reaction (PCR). A 14-kb clone was isolated from a flounder genomic library constructed in lambda FIXII. Of this clone, 8 kb was sequenced, including 3 kb of upstream sequence. The predicted amino acid sequence showed closest similarity to plaice CYP1A1 (98%). Gene structure conformed to the seven exons and six introns common to previous CYP1A sequences, but intron lengths were not conserved. Concensus sequences corresponding to xenobiotic and other response elements as well as TATA, CAAT and GC boxes were identified. Upstream sequence (3.5 kb) including the first exon and intron up to the putative start codon were amplified via PCR and inserted upstream of the luciferase gene in a pGL3 reporter gene construct. The HepG2 mammalian hepatoma cell line was transiently co-transfected with the flounder CYP1A reporter gene construct and the pRL-CMV internal control construct. The maximal induction upon exposure to 100 nM 3-MC was 4.4-fold in comparison with carrier-treated cells. Use of deletion constructs resulted in loss of inducibility. 相似文献
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The cytochrome P450 1 (CYP1) family has expanded with the addition of the CYP1B and CYP1C subfamilies. We recently identified a new CYP1 subfamily in zebrafish, CYP1D, with a single gene, CYP1D1. Here we examined sequences found in other fish genomes, i.e., stickleback (Gasterosteus aculeatus) and medaka (Oryzias latipes), for similarities among fish CYP1D1 genes. The full-length deduced amino acid sequences for CYP1D1 in these two species averaged about 43% identity to the CYP1As, but nearly 50% when sequence alignment ambiguities were masked. CYP1D1 has seven exons, similar in size and position to the exons in CYP1D1 and CYP1A in zebrafish. However, the intronic distances were substantially smaller in the medaka and stickleback. There also were differing numbers of putative xenobiotic response elements in the CYP1D1 of the various species. Whether the stickleback or medaka genes are inducible by aryl hydrocarbon receptor (AHR) agonists is yet to be determined. 相似文献
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用蔗糖密度梯度离心法分离纯化白斑综合症病毒（WSSV）。设计并合成引物，提取WSSV中国株的基因组DNA作为模板，通过PCR，扩增克隆出VP28基因，利用BamHI和EcoRI切点将VP28基因插入克隆载体pUC19 的多克隆位点上，得到VP28基因的重组克隆质粒，对其进行双向DNA测序。测序结果表明，该基因含有615个核苷酸，与GenBank中已有不同来源的WSSV的序列片段同源性为100％。利用BamHI切点将VP28的基因插入到穿梭表达载体启动子PpsbA的下游，EcoRI酶切鉴定，得到正向连接的可在蓝藻表达的重组穿梭表达载体，命名为pRL-VP28。 相似文献
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为了研究大菱鲆(Scophthalmus maximus)中趋化因子(Chenokine)的特征及其在大菱鲆免疫过程中发挥的作用,设计了本文中的实验。本实验从大菱鲆基因组和转录组数据库中鉴定了一个硬骨鱼特有的趋化因子CC亚家族成员—CCL34,并选取大菱鲆的8个健康组织以及2种细菌感染后的肠道和皮肤组织,采用实时荧光定量PCR(qRT-PCR)对其表达特征进行研究。序列分析结果显示该趋化因子全长mRNA包含1个327 bp的5’非编码区(UTR),1个246 bp的3’非编码区,和1个编码103个氨基酸残基长度为312 bp的开放阅读框(ORF)。此CCL34基因含有4个外显子和3个内含子。通过系统发育分析、共线性分析,将该大菱鲆趋化因子命名为CCL34。实时荧光定量PCR表明CCL34在大菱鲆健康组织中普遍表达,尤其在肾脏、肝脏、皮肤中有高水平表达。鳗弧菌(Vibrio anguillarum)和无乳链球菌(Streptococcus agalactiae)感染后,肠道组织CCL34基因表达量较对照组显著性上调(p<0.05),这表明CCL34可能在大菱鲆的肠道粘膜免疫中起重要作用。本研究为趋化因子家族功能的研究奠定了基础,为增强鱼类免疫力和抗病能力提供了理论依据。 相似文献
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CHU Wuying YU Lian QIAN Ronghu MENG Tao ZHOU Ruixue FU Guihong CHEN Ji ZHANG Jianshe 《海洋学报(英文版)》2008,27(5):126-133
One adult α-globin gene and one β-globin gene have been cloned from the large yellow croaker Pseudosciaena crocea. Linkage analysis indicated that the α- and β-globin genes were oriented head-to-head relative to each other. To identify the regulatory elements present in the intergenic and intragenic regions of the globin complex, the intergenic region alone or together with the β-globin gene first intron was cloned into the luciferase-reporter vector pGL3-Basic respectively, and the chimeric constructs were tran- siently transfected into Vero cells and primary fish erythrocytes. The intergenic region cannot support the high-level expression of luciferase. However, the promoter activity of the intergenic region was strongly stimulated by the positive regulatory elements （PRE） located in the β-globin gene intron 1. Thus, it is proposed that the intergenic promoters and intragenic PRE were necessary for the effective expression of the linked α- and β-globin genes. 相似文献
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采用透射电镜技术,研究了3种鲈形目鱼类褐菖鲉Sebastiscus marmoratus、黑鲷Sparus macrocephalus及平鲷Rhabdosargus sarba(Forskal)精子发生早期精原细胞、初级精母细胞及次级精母细胞的超显微结构变化。结果表明,3种鱼生精细胞形态结构及代谢活动变化主要体现在核的变化、线粒体数目与结构的变化及高尔基体和溶酶体的行为变化。3种鱼核的变化基本一致:细胞核由近圆形或椭圆形转变为多角形;核质逐渐浓缩;核仁从明显结构至完全消失。褐菖鲉精原细胞时期线粒体分布较多,并出现拟染色质与线粒体相粘附;而平鲷与黑鲷精原细胞时期的线粒体分布较少,无拟染色质。3种鱼高尔基体与溶酶体的变化也有相似性。黑鲷的初级精母细胞中还观察到典型的环层状的髓样小体的结构。 相似文献
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