中国兽医科学2012，Vol.42Issue(7)：719-724,6.
鸡β防御素2成熟肽基因在毕赤酵母中的表达及其抗菌活性分析
Expression of Gal-2 mature peptide gene in Pichia Pastoris and analysis of antimicrobial activity of the recombinant protein
摘要
Abstract
To construct the recombinant Pichia pastoris expressing chicken beta-defensin-2（Gal-2） and research the antimicrobial activity, the gene encoding Gal-2 mature peptide was cloned into an expression vector pPIC9K which was then transformed into P. pastoris GSl15 by electroporation. The positive clones of recombinant yeast were screened and induced by methanol. The expressed supernatant was detected by Tricine-SDS-PAGE and antibacterial experiment. Finally, the prevention effect of expressed supernatant was researched by the experiment of chicken infected by Escherichia coli CVCC1565. The result showed that the recombinant secretion expression plasmid pPIC9K-Gal-2 was successfully constructed and the 5.8 ku chicken Gal-2 protein was detected by Tricine-SDS-PAGE. The expressed supernatant showed an inhibi- ting effect on the growth of many bacteria including two avian pathogenic bacteria of Salmonella Pullorum S2 and birds pathogenic E. coli CVCC1565. In the experiment of chicken infected by E. coli CVCC1565, compared to injection of empty yeast expression supernatant group and PBS control group, the mortality of Gal-2 of different concentration expression supernatant group was significantly decreased（P〈0.01）. The prevention effect of Gl-2 expressed supernatant reached high level. The recombinant P. pastoris GSl15 was successfully constructed,which provided the basis for research and development of chicken defensin as new feed additives.关键词
鸡口防御素2/成熟肽/分泌表达/抗茵活性Key words
Gal-2/mature peptide/secretion expression/antimicrobial activity分类
医药卫生引用本文复制引用
王海霞,缪刘,祁克宗,涂健,刘红梅,彭开松..鸡β防御素2成熟肽基因在毕赤酵母中的表达及其抗菌活性分析[J].中国兽医科学,2012,42(7):719-724,6.基金项目
国家高技术研究发展计划（863）项目 （）
