伊氏锥虫HGPRT缺陷株和自然株的HGPRT基因的分离、克隆与比较

ISOLATION,CLONING AND COMPARISON OF HGPRT GENE OF NATURAL AND HGPRT - STRAINS OF TRYPANOSOMA EVANSI

参照布氏锥虫 HGPRT基因的核苷酸序列 ,设计一对引物 ,分别以伊氏锥虫 HGPRT缺陷株和自然株的总 RNA为模板进行 RT—PCR扩增 ,结果自然株出现 630 bp的 DNA条带 ,与设计大小一致 ,而缺陷株均为阴性 ,证明缺陷株的 HGPRT基因发生明显突变或缺失。自然株 HG-PRT基因经与 p GEM- T Easy Vector连接 ,大肠杆菌转化 。

A pair of PCR primers for Trypanosoma evansi HGPRT gene was designed according to the HGPRT gene sequence of Trypanosoma burcei .With the primers,HGPRT gene(about 630bp)of natural strain was successfully amplified from natural strain but not HGPRT - strain by RT PCR.That showed HGPRT gene is mutated or deficiency in the HGPRT - strain.The HGPRT gene was cloned into pGEM T Easy Vector and the reconbinant plasmid was identified by PCR amplification test and nucleotide sequencing.