抗菌肽天蚕素B突变体ABP-S1基因在E.coli中的融合表达  

Expression of a Mutant Cecropin B Lytic Peptide Analog ABP S1 in E.coli as a Fusion Protein

作　　者：卢强[1] 刘维全[1] 赵凤芹[1] 刘明远[2] 江禹[1] 殷震[1] 

机构地区：[1]解放军军需大学军事兽医研究所,吉林长春130062 [2]解放军军需大学动物科技系,吉林长春130062

出　　处：《中国兽医学报》2002年第3期241-243,共3页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目 ( 39870 5 5 2 )

摘　　要：利用绿色荧光蛋白 (GFP)基因与天蚕素 B突变体 ABP- S1基因的融合基因 ,构建了 2个原核表达载体 p Am GS1和 p Bm GS1,转化表达宿主菌 E.coli BL 2 1(DE3)和 E.coli BL 2 1(DE3) p L ys S。结果 ,p Am GS1未能得到转化子 ,而p Bm GS1的转化子高效表达了融合蛋白 ,经 IPTG诱导 ,SDS- PAGE检查 ,融合表达产物最高可占菌体总蛋白的49.45 %。表达产物经包涵体复性 ,柱层析初步纯化 ,通过平板抑菌试验 ,对大肠杆菌、金黄色葡萄球菌、铜绿假单胞菌、变形杆菌以及鱼类重要的致病菌嗜水气单胞菌等多种革兰氏阳性。The ABP S1 gene was fused with green fluorescent protein(GFP) gene which the stop codon TAA was replaced by restriction enzyme BglⅡ cut sequence by PCR, the fusion gene was named mGS1. Two prokaryotic expression plasmids pAmGS1 and pBmGS1 were constructed by cloning the fusion gene mGS1 into pET20b(+) and pET28b(+),respectively. E.coli competent host BL21(DE3) and BL21(DE3)plysS were transformed with the two expression plasmids, the pAmGS1 failed to get any transformants, but the pBmGS1 was successfully transformed. Both BL21(DE3) and BL21(DE3)plysS recombinant bacteria could express the mGS1 gene at high level, amounts to 49.45% of the total protein of the IPTG induced recombinant bacteria after 4.5 hours. The purified recombinant peptide showed the antibacterial activities upon several bacteria include E.coli, Staphylococcus aureus, Aeromonos hydrophlia, Salmonella spp etc, but did not show any antibacterial activities against Helicobacter Pylori.

关 键 词：抗菌肽 原核表达 绿色荧光蛋白 抗菌活性 大肠杆菌 天蚕素B 突变体 ABP-S1 E.COLI 基因表达 

分 类 号：Q786[生物学—分子生物学]