文章摘要
范武江,李思发,王兵,孟庆辉.尼罗罗非鱼、萨罗罗非鱼及其杂交子代胰岛素样生长因子(IGF-Ⅰb)基因3′cDNA末端克隆及序列分析[J].水产学报,2010,34(4):489~499
尼罗罗非鱼、萨罗罗非鱼及其杂交子代胰岛素样生长因子(IGF-Ⅰb)基因3′cDNA末端克隆及序列分析
Cloning and sequencing of IGF-Ⅰb 3′cDNA in Oreochromis niloticus, Sarotherodon melanotheron and their hybrid
投稿时间:2009-09-27  修订日期:2009-12-22
DOI:10.3721/SP.J.1231.2010.06625
中文关键词: 尼罗罗非鱼  萨罗罗非鱼  杂交子代  胰岛素样生长因子-Ⅰb  cDNA  克隆
英文关键词: Oreochromis niloticus, Sarotherodon melanotheron, Oreochromis niloticusх Sarotherodon melanotheron ,IGF-I,cDNA clone
基金项目:国家重点基础研究发展计划
作者单位E-mail
范武江 上海海洋大学 fanwujiang@hotmail.com 
李思发 上海海洋大学 sfli@shfu.edu.cn 
王兵 上海海洋大学农业部水产种质资源与利用重点开放实验室  
孟庆辉 上海海洋大学  
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中文摘要:
      为探讨尼罗罗非鱼、萨罗罗非鱼及其杂交子代(Oreochromis niloticus♀× Sarotherodon melanotheron)IGF-Ⅰ基因结构差异,研究IGF-Ⅰ基因与此3种基因型罗非鱼耐盐性能差异之间的关系,通过3′ RACE的方法,从鳃的总RNA分别获得其IGF-Ⅰb基因的部分cDNA序列,长度分别为1 076 bp、1 075 bp和1 079 bp,包含完整的开放阅读框(ORF)546 bp、一个终止密码及含polyA信号的3′UTR;ORF编码182个氨基酸,包括信号肽(44aa),成熟肽B区(29aa)、C区(10aa)、A区(21aa)、D区(8aa)及编码70aa 的E区,二级结构为混合类型;与其他脊椎动物IGF-Ⅰb序列的序列相似度达75.8%~100%;成熟肽A、B区高度保守,C区第82位后缺失2aa, E区第131位和159位分别缺失3aa和1aa; 萨罗罗非鱼在E133位发生Ala/Pro氨基酸残基替换,与耐盐性强的莫桑比克罗非鱼和画眉罗非鱼一致,推测IGF-Ⅰb基因E区选择性剪切与3种罗非鱼耐盐性能差异有关。
英文摘要:
      Aiming at exploring the sequence differences of IGF Ⅰ gene of Nile tilapia (Oreochromis niloticus NewGifts,ON), black chin tilapia (Sarotherodon melanotheron,SM) and their First Generation of Hybrid (HB), investigating the relationships between the great differences of salt resistant property among the three genotypes, we cloned partial cDNA fragments of insulin like growth factor Ⅰ(IGF-Ⅰ) from gill extracts through rapid amplification of 3′ cDNA ends(3′RACE)methods. The results demonstrated to be the IGF-Ⅰb gene cDNAs with length of 1 076 bp,1 075 bp and 1 079 bp respectively and comprising of a stop codon, a 546 bp open reading frame (ORF) encoding a 182 amino acid peptides(aa), and 3′ untranslated regions(3′ UTR). The deduced amino acid sequence of IGF-Ⅰb precursor is composed of signal peptide, mature peptide(including conserved B, C, A and D domains consisting of 29 aa,10 aa,21 aa and 8 aa respectively)and E domain of 70aa. Results of the predicted secondary structures were typed as “mixed patterns”. The results of multiple alignments showed highly homologous with 75.8% to 100% amino acid identity. The B and A domains are highly conserved in all of the allied species. There are 2aa deletion between No.82 and 83 sites in C domain, 3aa and 1aa deletion next to NO.131 and No.159 site, respectively. Ala / Pro substitution occurred at No.133 of E domain of black chin tilapia compared with ON and HB, which is identical to Oreochromis mossambicus and Oreochromis urolepis hornorum. We presumed that the differences of salt resistant property of ON, SM and HB may be due to the alterative splicing of the IGF-Ⅰ E domain.
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