扩展莫尼茨绦虫β微管蛋白基因的克隆及序列分析  

Cloning and Sequence Analysis of β-tubulin from Moniezia expansa

作　　者：夏党荣[1,2] 王涛[1,2] 薄新文[1] 马勋[2] 何延华[1] 康立超[1] 王新华[1] 

机构地区：[1]新疆农垦科学院/新疆生产建设兵团绵羊繁育生物技术重点实验室,新疆石河子832000 [2]石河子大学动物科技学院,新疆河子832000

出　　处：《新疆农业科学》2012年第2期324-329,共6页Xinjiang Agricultural Sciences

基　　金：国家重点基础研究发展计划("973")前期研究专项(2006CB708512);国家绒毛用羊产业技术体系建设专项(CARS-40-11);中国农业科学院兰州兽医研究所国家重点实验室开放课题(SKLVEB2011KFKT008);新疆兵团杰出青年科学基金(2011CD005)

摘　　要：【目的】从扩展莫尼茨绦虫中克隆β微管蛋白基因,进行序列测定和生物信息学分析,为进一步研究该基因的功能奠定基础。【方法】构建扩展莫尼茨绦虫成虫cDNA文库,随机挑取重组阳性克隆进行测序,对部分序列进行引物步移法测序,获得其全长cDNA序列;采用生物信息学等技术对该cDNA序列进行开放阅读框(ORF)的寻找、编码氨基酸的推导、核苷酸和氨基酸同源性比较以及蛋白二级结构的初步预测。【结果】获得了1个扩展莫尼茨绦虫新基因——β微管蛋白基因,全长1 571 bp,编码444个氨基酸,与日本血吸虫的β微管蛋白氨基酸序列具有78.6%的同源性。编码蛋白的理论分子量为49.6 ku,等电点为4.96;1～4位氨基酸MREI为β微管蛋白转录后调控信号,140～146位GGGTGAG存在一个微管蛋白标志信号片段,在99～106位和391～410位存在两个典型的β微管蛋白保守区;亚细胞定位分析其为细胞的骨架蛋白。【结论】获得了扩展莫尼茨绦虫β微管蛋白基因的全长cDNA序列,为该基因功能的实验性鉴定工作奠定基础。[ Objective ] The purpose of this project was to clone β - tubulin gene from Moniezia expansa （ M. expansa） cDNA library and then sequence and analyze it from the perspective of bioinformatics in order to provide a foundation for further research. [ Method ] A cDNA library was constructed from M. expartsa. Clones were selected randomly from the eDNA library and were sequenced by using the method of expression sequence tags （ESTs）. Novel genes were acquired by primer - walking. The eDNA sequence encoding M. ex- pansa ~ -tubulin was analyzed, including searching the ORF, translating the nucleotide to protein sequence, similarity searches and secondary structure predication. [ Result ] β -tubulin genes, 1 571 bp and coding for 444 amino acids was got. The β - tubulin of M. expansa and Schistosoma japonicum were homologous with an identity of 78.6 %. The molecular weight of the protein was predicted to be 49.69 Ku, and the academic pI was 4.96. The MREI domain, a β -tubulin mRNA autoregulation signal, was found at the N -terminus. There was a tubulin signature sequence GGGTGAG at the sites 140 - 146 of the deduced amino sequence and two conserved amino acid sequence of β - tubulin at the sites 99 - 106 and 391 -410. It is a kind of cytoskel- etal protein at subcellular location. [ Conclusion ] The full - length cDNA encoding M. expansa β - tubulin was obtained, which laid the foundation for further functional study of this gene.

关 键 词：扩展莫尼茨绦虫 β微管蛋白 克隆 序列分析 

分 类 号：S852.6[农业科学—基础兽医学]