银鲫两个蛋白合成相关基因全长cDNA的克隆及其特征分析  

CLONING AND CHARACTERIZATION OF FULL LENGTH cDNAs OF TWO GENES RELATED TO PROTEIN BIOSYNTHESIS IN GYNOGENETIC GIBEL CARP

作　　者：刘军[1] 石耀华[1] 尹隽[1] 桂建芳[1] 

机构地区：[1]中国科学院水生生物研究所

出　　处：《水生生物学报》2003年第5期512-520,共9页Acta Hydrobiologica Sinica

基　　金：国家自然科学基金重点项目 ( 3 0 13 0 2 40 );中国科学院知识创新工程重要方向项目 (KSCX2-SW-3 0 3 );中国科学院水生生物研究所知识创新工程领域前沿项目 ( 2 2 0 3 0 9)资助

摘　　要：通过构建雌核发育银鲫心跳期SMARTcDNA质粒文库并从文库中随机挑选克隆测序 ,克隆得到银鲫翻译起始因子 3亚单位 2 (GTIF3 S2 )和翻译延伸因子 1亚单位α(GEF 1α)基因全长cDNA。银鲫翻译起始因子 3亚单位 2基因cDNA全长 12 80bp ,开放阅读框位于 117— 10 91bp之间 ,编码 32 5个氨基酸。其推断的氨基酸序列存在三个WD结构域。该基因在鱼类中为首次报道。银鲫翻译延伸因子 1亚单位alpha基因cDNA全长 1784bp ,开放阅读框位于82— 14 6 7bp之间 ,编码 4 6 2个氨基酸。RT PCR表明 ,这两个基因在成熟卵母细胞和胚胎发育早期可以检测到少量的转录产物 ,在胚胎发育期间从原肠期开始转录 ,并随着发育进程逐渐增强。成鱼组织中除精巢表达较弱外 ,其他组织都表达较强。同源分析比较表明 ,TIF3 S2和EF 1α在物种进化过程中具有高度的进化保守性 ,在物种间的同源性很高。因此 ,作者认为 ,这两个基因是研究物种间系统发育的优良对象。The SMART cDNA plasmid library was constructed from heartbeat stage embryos of gibel carp(Carassius auratus gibelio). Randomly selected clones from the library were sequenced and their homologies were analyzed. The full length cDNAs of two genes related to protein biosynthesis were obtained from the sequenced clones. One gene,gibel carp translation initiation factor 3 subunit 2,was named GTIF3-S2. It is 1280 bp in length and has an open reading frame of 975 nucleotide acids for encoding 325 amino acids. The deduced amino acid sequence has three WD domains. The gibel carp GTIF3-S2 was first reported fish translation initiation factor 3 subunit 2. Another gene,gibel carp translation elongation factor 1α, was named GEF-1α. It is 1784 bp in length and has an open reading frame of 1386 nucleotide acids encoding 462 amino acids. The open reading frame is between 82—1467 bp. RT-PCR analysis showed that the maternal mRNAs of the two genes were detected in matured eggs and in early developmental embryos. In embryonic development, the two genes began to transcribe from gastrula stage, and their expressions became stronger as the embryos developed. The two genes were detected to express strongly in most of the examined tissues of gibel carp,but weaker expressions were found in testis. Homology analysis indicated that there exists high evolutionary conservation in gibel carp GTIF3-S2 and GEF-1α. among different species, from animals to plants. Therefore, we conclude that gibel carp GTIF3-S2 and GEF-1α. are two excellent target genes for studies on evolutionary genetics and systematics among species.

关 键 词：银鲫 起始因子 延伸因子 基因克隆 表达特征 CDNA 

分 类 号：Q959.468[生物学—动物学] S965.117[农业科学—水产养殖]