河南HIV感染者的HIV-1 B型株tat基因的克隆及序列分析

Cloning of tat Gene of a HIV-1 B Strain From Henan Province and Homologus Analysis of its Encoding Product

克隆河南人免疫缺陷病毒(HIV)感染者HIV-1B型株tat基因完整编码框序列,并分析比较其编码产物的序列结构特点。使用重叠PCR技术,从河南省1名HIV-1感染者外周血标本中扩增出tat基因第一和第二外显子并重组为完整的tat基因序列。获得的HIV-1B病毒株tat基因,第一外显子为263bp,第二外显子为214bp。将该基因编码产物与其他HIV-1株Tat蛋白经DNA软件编辑并翻译成蛋白质,使用ClustalX1.81进行多序列对比分析发现,第一外显子编码产物的3个保守区域的氨基酸组成大致相同,只有少数氨基酸存在差异。由于Tat蛋白不同病毒株间有高度保守的Cys富集区、核心区和碱性氨基酸富集区,tat基因的克隆为研究其功能并以其为靶点设计和筛选抗艾滋病的药物奠定了基础。

Clone the full-length sequence of tat gene of a HIV-1 B strain from Henan Province of China, and compare the similarity of its encoding product with known tat protein. By means of overlap PCR, the full-length sequence of HIV-1 tat gene was amplified from a HIV-1 infected patient's leucopheresis peripheral blood cells, the homologous analysis was proformed by Clustal X1.81. The full-length tat gene was obtained and sequenced. Exon 1 and exon 2 is 263 bp and 214 bp respectively. By comparing its encoding product with the reported HIV-1 Tat protein, we found that the composition of the main functional amino acid resudis is similar among these Tat proteins from different strains. However, a few differences of amino acid residues were found at the three conservative functional domains: cysteine-rich domain, core domain and basic amino acid domain. Tat protein play a crucial role in the process of HIV infection and pathogenesis. The cloned tat gene can be used to study the function of tat protein and screen its antagonists for AIDS therapy.