中国动物传染病学报2012，Vol.20Issue(6)：29-35,7.
热敏肠毒素促进产肠毒素大肠杆菌对小肠上皮细胞系IPEC-J2的黏附的初步研究
HEAT-LABILE ENTEROTOXIN ENHANCES ETEC ADHERENCE TO PORCINE SMALL INTESTINE EPITHELIAL CELL LINE IPEC-J2
摘要
Abstract
The LT gene with size of 1.1 kb was amplified in PCR using the genomic DNA template of K88ac E.coli strain C83902.The PCR products were digested with the restriction enzymes at each end and then cloned into vector pACYC184.The recombinant LT plasmids were then constructed and confirmed by restriction endonuclease analysis.In addition,two LT variants(pACYC-LT72 and pACYC-LT192)were also constructed,each bearing mutations of amino acid residues in the A subunit(A72R and R192G) that rendered the toxin inactive.The LT expression was confirmed in GM1-ELISA.Porcine small intestine epithelial cell line IPEC-J2 with glycoprotein receptors for bacterial in vitro adhesion was prepared and used in the study.The data demonstrated that elaboration of LT promoted a significant increase in E.coli adherence and the ADP-ribosylation activity was necessary for this process.We also demonstrated that the enhancement of bacterial adherence was blocked by protein kinase A inhibitor Rp–cAMP,adenylate cyclase antagonist DDA and LT receptor GM1.关键词
热敏肠毒素/黏附/IPEC-J2Key words
K88ac E.coli/heat-labile toxin/bacterial adherence/porcine small intestine epithelial cell line分类
农业科技引用本文复制引用
郁磊,吴娟,朱国强..热敏肠毒素促进产肠毒素大肠杆菌对小肠上皮细胞系IPEC-J2的黏附的初步研究[J].中国动物传染病学报,2012,20(6):29-35,7.基金项目
国家自然科学基金 （）
江苏省属高校自然科学重大基础研究项目 （）
江苏高校优势学科建设工程资助项目 （）
教育部创新团队 （）
科技部转基因生物新品种培育重大专项 （）
