伊氏锥虫HGPRT缺陷株和自然株的HGPRT基因的分离、克隆与比较

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６３０ｂｐ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ＨＧＰＲＴ基因经与ｐＧＥＭ－Ｔ Ｅａｓｙ Ｖｅｃｔｏｒ连接，大肠杆菌转化，获得了阳性克隆。

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

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.