关键词: 艾滋病痴呆综合症；HIV-1反式激活因子；pGEXKG载体；基因表达

Abstract:

Objective    To explore variations of the HIV-1 trans-activator of transcription(HIV-1 tat) exon 1 gene from three different tissues of a patient who died of AIDS dementia complex（ADC）and express the Tat-GST protein in Escherichia coli BL21, for investigating the effect of the variation on neurotoxicity of Tat. Methods    The tat gene was amplified with PCR from genomic DNA which was extracted from three different tissues, spleen(SPL), meninges(MG) and basal ganglia(BG) of the patient, and the PCR products were cloned into the PGEM-T vector. Sequence results were analyzed with BioEdit. After being cut by BamHⅠ-and EcoRⅠ and reclaimed, the tat gene was cloned into the pGEXKG vector. The recombinant plasmid pGEX-KG-tat was transferred and expressed in E. coli BL21. Expressed products were identified by SDS-PAGE and Western blot. Results    The tat exon 1 gene was successfully cloned. The sequence analysis showed that variations of the HIV-1 tat gene extracted from peripheral and central nerve tissues were different. The recombinant plasmid pGEX-KG-tat was effectively transferred and expressed in E. coli BL21, and SDS-PAGE and Western blot analyses which showed that the expressed protein was Tat. Conclusion    The tat exon 1 gene from peripheral and central nerve tissues was cloned and variations  exist and the TatGST fusion protein is efficiently expressed in E.coli, which can contribute to further research on neurotoxicity of Tat.

Key words: AIDS dementia complex; HIV1 transactivator of transcription; pGEXKG vector; Gene expression