贵州白香猪Gadd45G的cDNA克隆与序列分析  

Cloning and sequence analysis of Gadd45G cDNA from Guizhou Bai Xiang Pig

作　　者：乜玉丽[1] 许厚强[1] 赵佳福[1] 张勇[1] 陈祥[1] 

机构地区：[1]高原山地动物遗传育种与繁殖省部共建教育部重点实验室/贵州大学贵州省动物遗传育种与繁殖重点实验室,贵州贵阳550025

出　　处：《广东农业科学》2012年第10期141-144,共4页Guangdong Agricultural Sciences

基　　金：转基因生物新品种培育科技重大专项子项目(2009ZX08009-139B)

摘　　要：采用试剂盒提取RNA和RT-PCR法获得贵州白香猪Gadd45G基因cDNA序列,构建了贵州白香猪Gadd45G基因的pMD19亚克隆载体,并对该重组质粒进行测序分析,为贵州白香猪作为模式动物提供理论基础,为构建Gadd45G基因真核表达载体和转基因猪研究奠定基础。PCR、双酶切鉴定结果表明,已成功克隆的贵州白香猪Gadd45G基因的cDNA序列长度为480 bp。测序结果显示,贵州白香猪Gadd45G基因cDNA序列与普通猪、人类、家鼠、牛的同源性分别为99.6%、89.6%、90.2%、91.2%,氨基酸同源性分别为98.8%、95.6%、95.0%、96.2%。序列分析表明,贵州白香猪Gadd45G基因编码序列与普通猪相比,有两处发生碱基突变,其中一处为错义突变(第385处的G→A突变)使氨基酸由丙氨酸突变成苏氨酸。In order to obtain the cDNA sequence of Gadd45G, the RNA kit and the RT-PCR were used to construct the Godd45G pMD19-T vector of Guizhou Bai Xiang pig. Furthermore, the sequence of this recombinant plasmid was analyzed, which could provide theoretical basis for Guizhou Bai Xiang pig as model organisms and lay a foundation for constructing the eukaryon expression vector of Gadd45G and researching on transgenic pig. The Gadd45G cDNA coding sequence with 480 bp was cloned from Guizhou Bai Xiang pig, which identified by PCR and double digestion test. Compared with the sequence of Sus scrofa, Homo sapiens, Mus mnsculns, and Bos taurus reported in GenBank, the homology analysis showed that the homology rate of the Gadd45G cDNA sequence were 99.6%, 89.6%, 90.2% and 91.2% respectively, and the homology rate of amino acid sequence were 98.8%, 95.6%, 95.0% and 96.2% respectively. Compared with Sus scrofa reported in GenBank, the sequence analysis showed that there were two base mutations, one was nonsense mutations, the other was missense mutation： G→A at the positions of 385, and this mutation resulted in an amino acid change： alanine→threonine.

关 键 词：贵州白香猪 Gadd45基因 pMD19-T载体 克隆 序列分析 

分 类 号：Q344.12[生物学—遗传学]