安徽农业大学学报2012，Vol.39Issue(4)：556-559,4.
纳豆激酶基因的克隆和高效表达
Cloning and high-efficiency expression of nattokinase gene
摘要
Abstract
In this study, the nattokinase gene with signal peptide, propeptide and mature peptide was cloned and expressed in Escherichia coli, and the induction and expression parameters were optimized. The gene was amplified with genomic DNA extracted from Bacillus subtillis as template by RT-PCR, digested and ligated with expression vector-pET28a to construct recombinant plasmid pET28a-NK which was transformed into E coli strain BL21 (DE3). Induction conditions such as time, temperature and IPTG concentration were studied, and the highest fibrinolytic activity was obtained and it could reach 81.73 U·mL-1 when recombinant strain BL21 (DE3)-NK was cultivated at 20℃ for 20 h with IPTG at 0.4 mmol·L-1. An expected protein band about 38 kDa was observed by SDS-PAGE.关键词
纳豆激酶/枯草芽孢杆菌/基因克隆/表达Key words
nattokinase/ Bacillus subtillis/ gene clone/ expression分类
生物科学引用本文复制引用
王伟,甘露,甘旭华,陈晓琳,倪敬田,唐欣昀..纳豆激酶基因的克隆和高效表达[J].安徽农业大学学报,2012,39(4):556-559,4.基金项目
安徽省教育厅自然科学研究重点项目(KJ2009A107)资助. （KJ2009A107）
