富甘氨酸果蝇抗菌肽在大肠杆菌中的直接表达和纯化  

Direct Expression and Purification of Glycine-rich Antibacterial Peptide of Drosophila in Escherichia coli

作　　者：冯志国[1,2] 陆婕[1] 郎君[1] 李燕娇[1] 陈正望[1] 

机构地区：[1]华中科技大学分子生物物理教育部重点实验室,武汉430074 [2]信阳师范学院生命科学学院,信阳464000

出　　处：《生物技术通报》2009年第3期88-90,共3页Biotechnology Bulletin

基　　金：国家自然科学基金(30370647)

摘　　要：利用RT-PCR技术从果蝇总RNA中克隆出富甘氨酸果蝇抗菌肽基因,将该基因插入原核表达载体pET-32a(+)的Nde I和Xho I酶切位点之间,然后用重组质粒转化大肠杆菌BL21(DE3)利用乳糖操纵子的葡萄糖效应IPTG诱导表达,经15%SDS-PAGE、Western blot分析,在大约8 kD处出现了含6×His标签的抗菌肽蛋白。再以Ni-N TA纯化树脂进行亲和层析、脱盐、冻干,成功制备了富甘氨酸果蝇抗菌肽蛋白。The gene of antibacterial peptide was amplified from Drosophila total P, NA using RT-PCR,and then inserted into Nde I and Xho I enzyme-cutting sites of pET-32a plasmid. The recombinant vectors were transformed into the competent cell E. coli BL 21. By using the glucose effect or catabolite,the antibacterial peptide was expressed in E. coil BL21 （DE3） by the induction of IPTG. By the analysis of 15% SDS-PAGE and Western blot,the antibacterial peptide was found at 8 kD. After the expressed product was purified by affinity binding chromatography with Ni-NTA, salt-out and freeze-dried procedure,the product was obtained.

关 键 词：果蝇 富甘氨酸抗菌肽 直接表达 纯化 葡萄糖效应 

分 类 号：Q78[生物学—分子生物学] S888.31[农业科学—特种经济动物饲养]