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Position: Home > Articles > Expression of p33 gene of Theileria sergenti in BCG Chinese Veterinary Science 2013 (11) 1180-1183

瑟氏泰勒虫p33基因在卡介苗中的表达

作  者:
许应天;张西臣;高旭;宋建臣
单  位:
延边大学
关键词:
瑟氏泰勒虫;p33基因;卡介苗;表达
摘  要:
为了构建表达瑟氏泰勒虫p33基因的重组卡介苗,以瑟氏泰勒虫基因组DNA为模板,采用PCR方法扩增瑟氏泰勒虫p33基因,将其克隆至pMD18-T载体中,进行PCR、EcoRⅠ+HindⅢ双酶切和测序鉴定。将p33基因亚克隆到大肠杆菌-分枝杆菌穿梭载体pMV361中,电穿孔转化卡介苗,经热诱导后对其表达产物进行SDS-PAGE和Western-blot分析。结果表明,瑟氏泰勒虫p33基因在重组卡介苗中成功获得了表达,表达产物可被瑟氏泰勒虫阳性血清所识别。本研究结果为重组卡介苗在瑟氏泰勒虫病免疫预防的研究奠定了基础。
译  名:
Expression of p33 gene of Theileria sergenti in BCG
作  者:
XU Ying-tian;ZHANG Xi-chen;GAO Xu;SONG Jian-chen;Agricultural College of Yanbian University;College of Animal Science and Veterinary Medicine,Jilin University;
关键词:
Theileria sergenti;;p33 gene;;BCG;;expression
摘  要:
In order to construct recombinant BCG expressing p33gene of Theileria sergenti,the p33gene was amplified by PCR using T.sergenti genomic DNA as template,cloned into the pMD18-T vector,and identified by PCR,EcoRⅠ+HindⅢrestriction enzyme digestion and sequencing.Then,the p33gene was subcloned into Escherichia coli-Mycobacteria shuttle vector pMV361,and transformed into BCG by electroporation.After heat induction,SDS-PAGE and Western-blotting was used to analyze the expressed protein.The results showed that the p33gene was expressed in the recombinant BCG successfully;the expressed protein could be recognized by positive serum against T.sergenti.It is the foundation to study recombinant BCG vaccines for immunoprophylaxis of theileriosis sergenti.

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