工业微生物2017，Vol.47Issue(4)：24-29,6.DOI:10.3969/j.issn.1001-6678.2017.04.004
嗜热古菌S-腺苷甲硫氨酸合成酶的序列分析、基因克隆与异源表达
Cloning, sequence analysis and expression of S-adenosyl-methionine synthetase gene from Methanopyrus sp.SNP6
摘要
Abstract
The genome of Methanopyrus sp.SNP6 was sequenced and the sam gene was PCR amplified followed by sequence analysis.The sam gene was cloned into the expression vector pET-28b(+),then transformed into Escherichia coli BL21 (DE3).The recombinant protein was induced for expression by IPTG and the expressed protein was detected by using SDS-PAGE.The results showed that sam gene was 1 194 bp in length,coding for 397 amino acids,its theoretical molecular weight was 44 086.4 Da and it was a homologous tetramer.Its protein sequence was conservative with some related archaea.SDS-PAGE showed that the recombinant protein could be expressed and the molecular weight was consistent with the expectation.The recombinant protein was intracellular.A part of expressed proteins were solvable and the other was inclusion bodies.In this study,the sam gene was successfully cloned and expressed.It would provide a foundation for further purification,structural and functional research of S-adenosylmethionine synthetase from Methanopyrus sp.SNP6.关键词
嗜热古菌/S-腺苷甲硫氨酸合成酶/异源表达/序列分析Key words
thermophilic archaea/S-adenosylmethionine synthetase/sequence analysis/prokaryotic expression引用本文复制引用
马云婷,裘娟萍,余志良..嗜热古菌S-腺苷甲硫氨酸合成酶的序列分析、基因克隆与异源表达[J].工业微生物,2017,47(4):24-29,6.基金项目
国家海洋经济创新发展区域示范项目(12PYY001SF08) （12PYY001SF08）
浙江省新苗人才计划项目(2015R403095). （2015R403095）
