牛源产Ⅱ型不耐热肠毒素大肠杆菌的分离鉴定及重组毒素制备

doi:10.11843/j.issn.0366-6964.2014.08.019

Abstract

Abstract:

Type Ⅱ heat-labile enterotoxin (LT-Ⅱ)-producing Escherichia coli is a common zoonotic pathogenic agent responsible for diarrhea.In order to improve the understanding of the epidemiological situation of LT-Ⅱ toxin in cattle，a total of 138 E.coli isolates from 2010 to 2012 were detected for LT-Ⅱ gene by PCR，and ten of these were sequenced.The polycistron ORF of LT-Ⅱc1 was cloned into the pET30a expression vector，translated into Rossatta competent cells，and induced for expressing the recombinant LT-Ⅱ toxin.The toxicity of the recombinant LT-Ⅱ toxin was preliminary tested by mouse Y1 adrenal cells.The results showed that eight samples were LT-Ⅱc1 producing E.coli，one was LT-Ⅱc4 producing E.coli and one was LT-Ⅱc6 producing E.coli from ten LT-Ⅱ positive.The results of SDS-PAGE and Western Blot showed that recombinant LT-Ⅱc1 toxin was prepared successfully，and the recombinant toxin had induced the apparent cytopathic effect for mouse Y1 adrenal cells，with the minimum lethal dose of 17.8 pg.The results presented here demonstrate that we isolated the LT-Ⅱ producing E.coli for the first time in China，and lay a preliminary foundation for further study of the LT-Ⅱ toxin.

CLC Number:

S852.612

YUAN Chao-wen，LIU Wen-xin，GUAN Wei-kun，MENG Xiang-qiu，TANG Jie，LI Xing-yue，ZHAO Zhi-teng，SHI Dong-fang. Isolation of Type Ⅱ Heat-labile Enterotoxin-producing Escherichia coli and Preparation of the Recombinant Toxin[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(8): 1336-1341.

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Isolation of Type Ⅱ Heat-labile Enterotoxin-producing Escherichia coli and Preparation of the Recombinant Toxin

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