恶性疟原虫海南株(FCC1)翻译控制肿瘤蛋白(TCTP)基因的克隆及序列测定

为了获得翻译控制肿瘤蛋白 (TCTP)基因 ,提取恶性疟原虫海南株 (FCC1)总RNA ,直接用总RNA反转录成单链DNA ,再用长距PCR方法扩增出双链cDNA .根据小鼠约氏疟原虫TCTP基因序列设计引物 ,以cDNA为模板合成了恶性疟原虫海南株TCTP基因 .以pMD18 T为载体 ,用大肠杆菌XL1 blue对基因克隆 .测序结果表明 ,此基因序列与小鼠约氏疟原虫TCTP基因序列有 85 %同源性 .由此基因序列推导出的TCTP氨基酸序列 ,与小鼠约氏疟原虫TCTP氨基酸序列有 88%同源性 .进入GenBank国际基因库 (美国 )检索 ,所克隆的基因与基因库中恶性疟原虫基因同源性为 97% ;由所克隆的基因序列推导的TCTP氨基酸序列 ,与国际基因库恶性疟原虫TCTP基因推导的TCTP氨基酸序列仅有 1个氨基酸差异 .恶性疟原虫海南株TCTP基因克隆为进一步研究奠定了基础

Cloning and Sequencing of the TCTP Gene of Plasmodium falciparum from Hainan(FCC1)

To obtain the translationally controlled tumor protein(TCTP)gene,the total RNA of Plasmodium falciparum from Hainan(FCC1) was extracted . The total RNA was directly used in single-stranded(ss) DNA synthetic reaction.Double-stranded(ds) cDNA was selectively amplified by PCR. According to the known TCTP sequence of Plasmodium yoelii(P.yoelii),a pair of primers were designed. TCTP gene was amplified by PCR. The gene was inserted into pMD-18-T vector and cloned in the Xl1-blue. The TCTP of FCC1 showed 85% homology at nucleotide sequence and 88% homology at the amino acid sequence to TCTP of P.yoelii.The TCTP of FCC1 and the TCTP of Plasmodium faciparum from GenBank exhibited 97% homology in nucleotide sequence and 99% homology in amino acid sequence.The results laid a foundation for further research.