林麝、马麝及梅花鹿活化素基因β_A亚基成熟肽序列的克隆和分析  

Molecular Cloning and Sequence Analysis of Activin Gene β_A Subunit Mature Peptides from Forest Musk Deer, Alpine Musk Deer and Spotted Deer

作　　者：邹方东[1] 张义正[1] 杨楠[1] 岳碧松[1] 

机构地区：[1]四川大学生命科学学院

出　　处：《动物学杂志》2004年第3期22-27,共6页Chinese Journal of Zoology

基　　金：科技部重大基础前期研究专项基金资助项目 (No .2 0 0 3CCA0 30 0 0 )

摘　　要：活化素 (Activin)对睾丸和卵巢产生多重调节作用 ,对动物的繁育非常关键。本文参考已克隆的其它物种的活化素基因 βA 亚基成熟肽序列 ,设计一对简并引物 ,通过PCR方法从林麝 (Moschusberezovskii)、马麝 (Moschuschrysogaster)和梅花鹿 (Cervusnippon)的基因组DNA中直接扩增目的基因片段。将目的片段分别克隆到pMD1 8 T载体中 ,并进行序列测定。DNA序列测定及分析表明 ,林麝、马麝和梅花鹿的活化素基因 βA 亚基成熟肽序列长 3 45bp ,三物种核苷酸序列同源性在 98%以上 ,氨基酸序列同源性在99%以上 ,核酸限制性酶切图谱也高度相似。将它们与GenBank中已公布的其它物种的活化素序列进行比较 ,发现该片段在不同进化程度的物种间高度保守。这是首次从麝科动物中成功克隆与生殖相关的核基因 。Activins play very important roles in regulating the physiology of ovary and testis, and are pivotal hormones to organism′s reproduction. The sequences of Activin gene β-A subunit mature peptides have been successfully amplified from Forest musk deer (Moschus berezovskii), Alpine musk deer (Moschus chrysogaster) and Spotted deer (Cervus nippon) genomic DNA by polymerase chain reaction (PCR) with a pair of degenerate primers. They were separately cloned to the vector pMD18-T and transformed into JM109 cells. Sequence analysis of Activin gene β A subunit mature peptides demonstrated that the length of the three target fragments was 345 bp, and there were about 98% DNA homology and 99% amino acid homology among these three species. GenBank blast search results and restriction enzyme maps revealed that sequences of Activin gene β-A subunit mature peptide of different species were highly conserved during the evolution process. This is the first report describing the cloning of a reproduction related gene from musk deer.

关 键 词：林麝 马麝 梅花鹿 Activin基因βA亚基成熟肽克隆 

分 类 号：Q754[生物学—分子生物学]