野猪肌细胞生成素基因的克隆与序列分析  

Cloning and sequence analysis of myogenin gene in wild boar

作　　者：徐怀亮[1] 姚永芳[1] 朱庆[1] 潘阳[2] 

机构地区：[1]四川农业大学动物科技学院,四川雅安625014 [2]昆明动物园,云南昆明650021

出　　处：《浙江大学学报（农业与生命科学版）》2010年第1期9-15,共7页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：中国博士后科学基金资助项目(2005037803);四川农业大学青年科技创新基金资助项目(00230500);四川农业大学"211工程"双支计划资助项目(01520500)

摘　　要：参考家猪(X89007)MyoG基因序列设计2对特异引物,用PCR方法首次从野猪基因组中扩增出2个大小分别约为1.6 kb和1.0 kb的DNA片段,其PCR产物经pMD-18T载体转化感受态DH5α株大肠杆菌,获得重组克隆子。DNA测序和序列拼接表明:野猪肌细胞生成素基因DNA序列长2 466bp,含完整的3个外显子和2个内含子,与家猪、牛、马、狗、小鼠和人的MyoG基因的cDNA序列同源性分别为99.8%、92.4%、92.7%、89.7%、90.8%和94%,其cDNA编码氨基酸序列与家猪、牛、马、狗、小鼠和人的同源性分别为100%、96.4%、95.9%、94.1%、96.4%、96.8%;对野猪MyoG基因组序列与9个家猪品种的相应同源序列进行比较,检出16个核苷酸变异位点,且其中有5个为家猪中不具有的新变异位点,其变异位点主要发生在内含子部分,尤其是内含子1中的变异位点比例最大(11个)。这些结果表明,野猪肌细胞生成素基因的编码序列在进化过程中是高度保守的,而内含子部分尤其是第1内含子具有丰富的序列多态性。对117个限制性酶切位点扫描分析发现,野猪MyoG基因核苷酸序列中含有78个酶切位点,其中有5个酶切位点包含变异位点,尤其是1 153位点的T突变成G所产生的SmaI(CCC/GGG)或XmaI(C/CCGGG)酶切位点为野猪所特有。所测DNA序列已提交到GenBank中,获得的序列号为:FJ356697。Based on myogenin （MyoG） gene sequence of domesticated pig （X89007）, two pairs of primer were designed and used to amplify an approximate 1.6 kb and 1.0 kb DNA fragment by PCR technique from genomie DNA sample of wild boar. The PCR produets were ligated into the pMD-18T vector, and then transformed into competent cells of E. coli DH5a. The DNA sequencing and combining result showed that the MyoG gene of wild boar was 2 466 bp （without primer sequence） in length and contained three complete exons and two introns. The homology analysis of the MyoG gene sequences by Clusta｜ W software indicated that, compared with domesticated pig, cattle, horse, dog, mouse and human, the eDNA sequence homologies of MyoG gene of wild boar were 99.8%, 92.4%, 92.7%, 89.7~, 90. 8%, 94.0%, respectively, and the homologies of amino acid sequence were 100%/00, 96.4%, 95.9%, 94.1%, 96.4%, 96.8%, respectively. In addition, a total of 16 nucleotide variable sites were found and five sites of which were specific to wild boar. Most of variable sites occurred in the introns, especially the first intron. Theses results demonstrated that all exons of MyoG gene were highly conservative, but its introns （especially intron 1） were more polymorphic in the process of wild boar evolution. Screened with 117 enzymes, 78 recognition sites were found, and five positions of them included the variable sites. Especially, Sinai （CCC/GGG） or XmaI（C/CCGGG） recognition sites at site 1 153 with T being mutated into G was specific in the wild boar. DNA sequences were deposited to GenBank, and obtained sequence No. was FJ356697.

关 键 词：野猪 肌细胞生成素 克隆 序列分析 

分 类 号：S821.81[农业科学—畜牧学] S813.1[农业科学—畜牧兽医]