牛瑟氏泰勒虫表面蛋白p33基因的真核表达

EUKARYOTIC EXPRESSION OF SURFACE PROTEIN P33 GENE OF THEILERIA SERGENTI FROM THE CATTLE

目的构建牛瑟氏泰勒虫表面蛋白p33基因的真核表达载体。方法根据已发表的牛瑟氏泰勒虫(Thei-leria sergenti)表面蛋白p33基因的核苷酸序列设计引物,应用PCR技术从牛瑟氏泰勒虫基因组DNA中扩增p33基因片段并克隆入pMD18-Tsimple载体。进一步将该基因插入到真核表达载体pVAXⅠ,转染Hela细胞后进行RT-PCR检测和IFA检测。结果牛瑟氏泰勒虫表面蛋白p3基因成连接到pVAXⅠ载体中,并在真核细胞中有效表达。结论本研究试验为今后该基因的进一步动物试验奠定了基础。

Objective In order to construct an eukaryotic expression vector of surface protein P33 gene of Theileria sergenti. Methods A pair of primers were designed and synthesized according to the sequence of P33 gene of Theileria sergenti reported in the GenBank. The P33 gene was amplified by PCR from the DNA genome of Theileria sergenti and was cloned into pMD18-T simple vector, then was inserted into eukaryotic expression vector pVAX I and transfected Hela cell. Results RT-PCR showed that P33 gene was effectively transcribed. IFA demonstrated that the gene was expressed in eukaryotic cell. Conclusion The results established a base for further study on the function of p33 gene from Theileria sergenti.