摘要
目的克隆格氏菌素T基因,在原核细胞中表达,并检测其抑菌活性。方法通过PCR技术,从3株L.gasseri北京分离株中特异性扩增格氏菌素T的成熟肽DNA编码片段(V11/gatA和V441/gatA),经TA克隆和序列测定后,进行同源比较分析。双酶切后的目的片段与表达载体pGEX6P1连接,得到高效融合表达质粒pGEXgatA/V11和pGEXgatA/V441,转化大肠杆菌BL21(DE3)PlysS,IPTG诱导表达,并检测表达产物的抑菌活性。结果目的蛋白以包涵体形式表达,表达量约29%。重组格氏菌素对于金黄色葡萄球菌、铜绿假单胞菌、痢疾志贺氏菌和伤寒沙门氏菌具有明显的抑制生长作用。结论所获得的重组格氏乳杆菌素具有明显的抑菌活性,有望成为新型抗菌素。
Objective To clone gassericin T gene, express in prokaryotic cells and identify the microbiostatic activity of expressed product. Methods Amplify the DNA fragments encoding the mature peptide of gassericin T from three L. gasseri strains isola- ted in Beijing by PCR. Analyze the homology of the amplified gene fragments V1-1/gatA and V44-1/gatA to the gassericin T gene of L. gassed strain isolated in Japan. Insert the two amplified gene fragments into expression vector pGEX-6P-1 to construct recombinant plasmids pGEX-gatA/V1-1 and pGEX-gatA/V44-1 respectively. Transform the constructed recombinant plasmids to E. coli BL21 （DE3） PlysS for expression under induction of IPTG. Results The homologies of Vl-1/gatA and V44-1/gatA to the gassericin T gene of L. gassed strain isolated in Japan were 98% and 99% respectively. The expressed product, existing in a form of inclusion body, contained about 29% of total somatic protein. Recombinant gassericln T showed significant inhibiting activity to the growth of Staphylococcus aureaus ,Pseudomonas aeruginosa,Shigella dysenteriae and Tyuphoidal salmonellosis. Conclusion The obtained recombinant gassericin T might be a novel antibiotic.
出处
《中国生物制品学杂志》
CAS
CSCD
2006年第3期244-248,共5页
Chinese Journal of Biologicals
