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目的在昆虫细胞中表达中国流行株 Ｂ亚型 ＨＩＶ－１ Ｇａｇ蛋白，制备重组 Ｇａｇ蛋白。方法将中国 株 Ｂ亚型 ＨＩＶ－１的 Ｇａｇ全基因序列，克隆到杆状病毒表达载体 ｐｆａｓｔｂａｃＩ中，构建了重组质粒 ｐｆａｓｔＧａｇ。经转座及平板 筛选，提取重组穿梭质粒 Ｂａｃｍｉｄ。经转染Ｓｆ９昆虫细胞后，获得了重组杆状病毒。用 ＳＤＳ－ＰＡＧＥ、Ｗｅｓｔｅｒｎ ｂｌｏｔ和间接免疫荧光实验分析表达的目的蛋白。结果重组病毒在昆虫细胞 Ｓｆ９中高效表达了中国株 Ｂ亚型 ＨＩＶ－１ Ｇａｇ蛋 白，表达的重组蛋白具有良好的抗原活性，可与ＨＩＶ－１标准阳性血清及ｐ２４单克隆抗体发生较强的免疫反应。结论 重组表达产物可用于艾滋病的免疫诊断和疫苗研究。
Objective To express HIV-1 Gag protein of Chinese subtype B strain in insect cells and prepare recombinant Gag protein. Methods The full-length Gag gene of HIV-1 of Chinese strain B was subcloned into the baculovirus expression vector pfastbacI to construct the recombinant plasmid pfastGag. After transposition and screening, the recombinant shuttle plasmid Bacmid was extracted. After transfected into Sf9 insect cells, a recombinant baculovirus was obtained. The expressed proteins were analyzed by SDS-PAGE, Western blot and indirect immunofluorescence assay. Results The recombinant virus highly expressed HIV-1 Gag protein of Chinese strain B in Sf9 insect cells. The expressed recombinant protein had good antigenic activity and could be immunized with HIV-1 positive serum and p24 monoclonal antibody reaction. Conclusion The recombinant expression product can be used in immunodiagnosis and vaccine research of AIDS.