中国兽医科学2018，Vol.48Issue(9)：1131-1138,8.DOI:10.16656/j.issn.1673-4696.2018.0162
柔嫩艾美耳球虫SSADH基因的克隆原核表达与序列分析
Cloning,prokaryotic expression and sequence analysis of Eimeria tenella SSADH gene
摘要
Abstract
To investigate the biological function of succinic semialdehyde dehydrogenase (SSADH) from Eimeria tenella,the specific primers were designed referring to the published sequence of E. tenella SSADH (EtSSADH). The gene was amplified by RT-PCR, and the PCR product was cloned into pGEX-4T-l vector for expression by induction of IPTG. Meanwhile,bioinformatics analysis was conducted for the amino acid sequence encoded by the gene. The results showed that the open reading frame was 1 896 bp in length,encoded a protein of 631 amino acids. The encoded protein mainly located in mitochondria, without signal peptide and transmembrane domain. Sequence alignment of EtSSADH with SSADH amino acids of different species showed that the encoded amino acid sequences of these genes were highly conserved. The recombinant protein with molecular weight of about 91. 18 ku was obtained. The successful cloning and expression of EtSSADH laid a foundation for the functional study of the enzyme.关键词
柔嫩艾美耳球虫/琥珀酸半醛脱氢酶/基因克隆/原核表达/序列分析Key words
Eimeria tenella/succinic semialdehyde dehydrogenase/gene cloning/prokaryotic expression/sequence analysis分类
农业科技引用本文复制引用
霍晨阳,李国清,廖申权,戚南山,李娟,吴彩艳,吕敏娜,林栩慧,胡俊菁,孙铭飞..柔嫩艾美耳球虫SSADH基因的克隆原核表达与序列分析[J].中国兽医科学,2018,48(9):1131-1138,8.基金项目
国家重点研发计划专项资助项目(2016YFD0501303) （2016YFD0501303）
广东省特支计划"科技创新青年拔尖人才"项目(2015TQ-01N407) （2015TQ-01N407）
广东省科技计划项目(2016B020202005,2016A040403079,2015A020209075,2015A020209073,2017A02-0225025,2015B070701015,2015B020230004) （2016B020202005,2016A040403079,2015A020209075,2015A020209073,2017A02-0225025,2015B070701015,2015B020230004）
