摘要
以往在对藤黄微球菌的研究中发现了促进复活因子(RPF),该因子可以有效促进休眠期的多种革兰氏阳性细菌复活和生长。生物信息学分析表明在多种革兰氏阳性细菌的基因组中均存在编码RPF样蛋白的基因。从GeneBank中获得了结核分枝杆菌、麻风分枝杆菌、谷氨酸棒杆菌和微链霉菌等革兰氏阳性细菌的多种RPF样蛋白相关信息,通过同源性分析发现这些蛋白均含有一个由75个氨基酸残基构成的高度保守序列,即RPF作用域。进一步对结核分枝杆菌H37Rv株的Rv1009进行了分析,并用PCR法克隆了其编码基因全长,定向克隆至pGEX 4T-2,在大肠杆菌BL21(DE3)中获得了Rv1009-GST融合蛋白的高效表达,为深入探讨其生物学功能奠定了基础。
Resuscitation-promoting factor(RPF) was identified in the research on Micrococcus luteus and it is confirmed that RPF can resuscitate the dormant cells and promote the growth of Gram-positive bacteria. There are many genes coding RPF-like proteins in the genomes of some Gram-positive bacteria. The information of different RPF-like proteins of Micrococcus luteus, Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium leprae, Strepto-myces coelicolor, Streptomyces avermitilis and Corynebacterium glutamicum were obtained from GeneBank. A RPF domain composed of 75aa was identified by sequence alignments. We further analysed the rv1009 coding product of Mycobacterium tuberculosis H37Rv, cloned the full length gene by PCR, then cloned into the vector of pGEX 4T-2 and high level expression of Rv1009-GST fusion protein is achieved in BL(21) bacteria.
出处
《生物技术通讯》
CAS
2003年第6期557-559,共3页
Letters in Biotechnology
关键词
结核分枝杆菌
促进复活因子
克隆
表达
Mycobacterium tuberculosis
resuscitation-promoting factor
cloned
expression
