伯克霍尔德菌假鼻疽菌素BipD蛋白的生产与纯化。

Production and purification of Burkholderia pseudomallei BipD protein.

作者信息

Visutthi Monton, Jitsurong Siroj, Chotigeat Wilaiwan

机构信息

Department of Biochemistry, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, Thailand.

出版信息

Southeast Asian J Trop Med Public Health. 2008 Jan;39(1):109-14.

PMID:18567449

Abstract

Type III secretion system (TTSS) clusters contain virulent genes of both animal and plant Gram-negative bacteria. The full length (933 bp) gene of bipD, a member of TTSS cluster of Burkholderia pseudomallei, was isolated by PCR amplification from B. pseudomallei DNA and cloned into pGEX 4T-1. GST-BipD fusion protein and BipD protein obtained by removal of GST using thrombin were employed to detect the presence of B. pseudomallei antibodies in sera obtained from melioidosis and non-melioidosis patients by immunoblotting. Sensitivity and specificity of BipD protein was 100% and 91.1%, respectively, whereas that of fusion protein was 77.8% and 90.0%, respectively.

摘要

III型分泌系统（TTSS）基因簇包含动物和植物革兰氏阴性菌的致病基因。通过PCR扩增从类鼻疽伯克霍尔德菌DNA中分离出类鼻疽伯克霍尔德菌TTSS基因簇成员bipD的全长基因（933 bp），并将其克隆到pGEX 4T-1中。利用凝血酶去除谷胱甘肽S-转移酶（GST）后获得的GST-BipD融合蛋白和BipD蛋白，通过免疫印迹法检测类鼻疽病和非类鼻疽病患者血清中类鼻疽伯克霍尔德菌抗体的存在。BipD蛋白的敏感性和特异性分别为100%和91.1%，而融合蛋白的敏感性和特异性分别为77.8%和90.0%。