恶性疟原虫云南株富含组氨酸蛋白II部分基因的克隆和序列分析

CLONING AND SEQUENCE ANALYSIS OF HISTIDINE RICH PROTEIN Ⅱ GENE FRAGMENT OF PLASMODIUM FALCIPARUM YUNNAN STRAIN

目的：测定恶性疟原虫云南株富含组氨酸蛋白 Ｉ Ｉ部分序列，了解该虫株与其它虫株 Ｈ Ｒ Ｐ Ｉ Ｉ基因序列的差异。方法：采用 Ｐ Ｃ Ｒ 方法特异性扩增 Ｈ Ｒ Ｐ Ｉ Ｉ基因片段，并将该基因片段克隆于 Ｍ １３ 载体，用双脱氧链末端终止法进行测序，应用 Ｐ Ｃ Ｇ Ｅ Ｎ Ｅ软件对不同虫株恶性疟原虫 Ｈ Ｒ Ｐ Ｉ Ｉ进行基因序列分析。结果：我国云南株恶性疟原虫与 ７ Ｇ８ 及 Ｄ１０ 虫株 Ｈ Ｒ Ｐ Ｉ Ｉ基因有不同程度的差异，３ 虫株间 Ｈ Ｒ Ｐ Ｉ Ｉ氨基酸序列同源性为７０．３％ ，核苷酸序列同源性为 ６８．６％ 。结论：云南株与７ Ｇ８ 及 Ｄ１０ 虫株 Ｈ Ｒ Ｐ Ｉ Ｉ基因存在差异。

AIM:To determine the nucleotide sequence of the partial gene of the histidine rich proteinⅡ (HRPⅡ) the Plasmodium falciparum PFD 3/YN and find out the differences of the HRPⅡ gene sequence between this isolate and other isolates.METHODS:The histidine rich protein Ⅱ gene fragment was amplified by polymerase chain reaction and cloned into M13 bacteriophage.M13 HRPⅡ single strand DNAs of three positive clones were extracted, respectively.The nucleotide sequence of the HRPⅡ gene fragment was determined by the dideoxy chain termination method.Using PCGENE software to compare and analyze the HRPⅡ gene sequence among the different isolates.RESULTS: Different degrees of diversity of the HRPⅡgene sequences were found among Plasmodium falciparum PFD 3/YN(from China)and two other isolates(7G8 from Brazil and D10 from Gambia).The HRPⅡ in the three isolates exhibited 70.3% homology in amino acid sequences and 68.6% homology in the nucleic acid sequences.CONCLUSION:There were differences in HRPⅡ gene sequence among the Plasmodium falciparum PFD 3/YN and two other isolates(7G8 and D10).