作　　者： ; ; ; ; ;

机构地区： 广西科学院

出　　处： 《广西农业生物科学》 2006年第4期285-289,共5页

摘　　要： 丙酮酸甲酸裂解酶是肠道细菌在厌氧代谢中十分关键的酶,丙酮酸甲酸裂解酶激活因子(pyruvateform ate lyase activator,PFL-A)在功能上具有重要的作用。为进一步研究PFL-A的激活机理,以大肠杆菌K-12的基因组为模板,通过G enB ank上公布的序列设计引物,扩增出目的基因,克隆到pM D 18-T载体,经测序,所扩增出的基因与p f l-act基因具有99%的同源性。将p f l-act连接到高效表达载体pET-22b(+)中,经异丙基硫代-β-D-半乳糖苷(IPTG)诱导,结果发现,p f l-act以包涵体形式表达。改变诱导剂、诱导剂量或培养温度,对包涵体的形成均没有明显的影响。 Pyruvate formate lyase （PFL） is a key enzyme acting in anaerobic metabolic pathway in enteric bacteria. As activator of PFL, pyruvate formate lyase activator （PFL-A） shows important effect in the function. For the further research in the activating mechanism, pfl-act was amplified with the genomic DNA of Escherichia coli using the primers designed by the sequence from GenBank, ligated with pMD18-T vector, and sequenced. The cloning pfl-act nucleotide had 99% identity with the objective gene. The segment of pfl-act was ligated with the high expressing vector pET-22b （＋）, induced by IPTC. The result showed that pfl-act was expressed by inclusion body. The inclusion body was not obviously changed by optimized the amount of inducer, the inducer or the temperature of cultivating.

关 键 词： 丙酮酸甲酸裂解酶激活因子 包涵体 高效表达

领　　域： [生物学]