重组鸡β-防御素在毕赤酵母中的表达及其抑菌活性  

Expression of chicken β-defensin-3 in Pichia pastoris and its antimicrobial activity

作　　者：万雪[1] 赵建增 高英杰[3] 宁官宝[4] 邢海云[2] 乔彦良[1] 李朝阳[1] 

机构地区：[1]山东信得药业集团公司 [2]北京大北农集团动物医学研究中心,北京100080 [3]吉林大学畜牧兽医学院,吉林长春130062 [4]山西农业大学动物科技学院,山西太谷030801

出　　处：《中国兽医学报》2008年第6期637-640,共4页Chinese Journal of Veterinary Science

基　　金：山东省科技发展计划资助项目(031030101)

摘　　要：将鸡β-防御素(Gal-3)基因片段插入到酵母表达载体pPIC9k,在毕赤酵母GS115中进行表达,并研究了重组Gal-3的诱导、表达规律和抗菌谱。结果发现,重组菌株在接受诱导培养24h后开始有明显的Gal-3表达,诱导72h时达到表达高峰,至少持续表达96h以上。Tricine-SDS PAGE电泳发现,在约6000位置出现预期的蛋白质条带,约占分泌总蛋白的12.6%。证实,重组Gal-3能完全抑制金黄色葡萄球菌、铜绿假单胞菌、志贺痢疾杆菌、普通变形杆菌、伤寒沙门菌、化脓性链球菌、大肠杆菌、肠炎沙门菌等细菌的生长,其中大肠杆菌和肠炎沙门菌是对重组Gal-3最敏感的菌株。The chicken β-defensin(Gallinacin-3)cDNA fragment was amplified by reverse transcription-polymerase chain reaction(RT-PCR)from the chicken.The RT-PCR products were inserted into the pGEM-T easy cloning vector,and then was sequenced.The result showed that the obtained 240 bp DNA fragment is identical to the Gal-3 cDNA sequence registered in GenBank.Then the fragment of mature Gallinacin-3 gene was inserted into the pPIC9k expression vector and pPIC9k-gal-3 was acquired.Then the recombinant plasmid was electr...

关 键 词：鸡 Β-防御素 毕赤酵母细胞 表达 抑菌活性 

分 类 号：S852.5[农业科学—基础兽医学]