鸡胚成纤维细胞ATP合成酶β亚基基因的克隆与测序  

Cloning and Sequencing of the ATP SynthaseβSubunit Gene of Chicken Embryo Fibroblast Cells

作　　者：吾鲁木汗·那孜尔别克 黄菊英 恩特马克·布拉提白 Wulumuhan Nazierbieke;Huang Juying;Entomack Borrathybay(College of Biology and Geography Sciences,Yili Normal University,Yining,Xinjiang 835000,China)

机构地区：[1]伊犁师范大学生物与地理科学学院,新疆伊宁835000

出　　处：《伊犁师范学院学报（自然科学版）》2020年第2期42-47,共6页Journal of Yili Normal University：Natural Science Edition

基　　金：国家自然科学基金项目(31360613).

摘　　要：用MEM培养基对CEF细胞进行培养,采用RNA提取试剂盒提取CEF细胞总RNA,通过RT-PCR技术扩增编码ATP合成酶β亚基的ATP5B基因片段,将其克隆至pMD18-T载体上,转化大肠杆菌DH5α感受态细胞,PCR技术鉴定重组载体后,测定ATP5B基因的核苷酸序列.结果表明,CEF细胞ATP5B基因长度为1401 bp,编码466氨基酸残基的多肽,与在GenBank数据库中已报道不同鸟类ATP5B基因序列的同源性在86.91%~99.64%之间.克隆的CEF细胞ATP5B基因,为进一步开展ATP5B基因的表达、纯化及其致病机理研究奠定了基础.The gene encodingβsubunit of ATP synthase(ATP5B)was amplified by RT-PCR with specific primers from total RNA of chicken embryo fibroblast(CEF)cells.The PCR products was cloned into the pMDTM18-T vector and transformed into competent cell of Escherichia coli DH5α,and the recombinants was screened by the blue-white spot assay and PCR.The sequence analysis showed that the coding region of the ATP5B gene of CEF cells was 1401 bp in length,and homology of ATP5B genes between the CEF cells and the previously reported different poultries in GenBank was 86.91 to 99.64%.In conclusion,the cloned gene ATP5B of CEF cells can be used for further study of the expression,purification and pathogenesis of ATP synthaseβsubunit.

关 键 词：CEF细胞 ATP合成酶β亚基 基因克隆 DNA测序 

分 类 号：Q786[生物学—分子生物学]