HIV衣壳蛋白P24与TRIM22在HEK293T细胞中的表达及共定位

Expressions and co-localization of HIV capsid protein p24 and TRIM22 in HEK293T cells

目的观察人类免疫缺陷病毒(HIV)衣壳蛋白P24与三基序蛋白22(TRIM22)在HEK293T细胞中的共定位情况。方法以慢病毒载体p LP1为模板,通过PCR法扩增p24编码序列,克隆至p DsRed-Monomer-N1真核表达载体。经菌落PCR、双酶切及DNA测序进行鉴定。将p DsRed-Monomer-N1-p24与p EGFP-N3-TRIM22载体共同转染HEK293T细胞,用荧光显微镜观察p24-DsRed-Monomer和TRIM22-EGFP的表达及共定位情况。结果经菌落PCR、双酶切及测序鉴定,成功构建p DsRedMonomer-N1-p24真核表达载体。通过荧光显微镜检测发现,p24-DsRed-Monomer和TRIM22-EGFP在HEK293T细胞中存在共定位关系。结论 HIV衣壳蛋白P24与TRIM22存在共定位。

Objective To observe the co-localization of human immunodeficiency virus（HIV） capsid protein p24 and tripartite motif containing 22（ TRIm^22） in HEK293 T cells. Methods The retroviral packaging vector p LP1 was used as the template of p24. After the amplification by PCR,the sequence of p24 was cloned into the eukaryotic expression vector p DsRed-Monomer-N1. The recombinant vector was confirmed by colony PCR,double restriction enzyme digestion and DNA sequencing. HEK293 T cells were co-transfected with the vector p DsRed-Monomer-N1-p24 together with p EGFP-N3-TRIM22.After 24 hours, the co-localization of p24-DsRed-Monomer and TRIM22-EGFP was detected under a fluorescence microscope. Results Colony PCR,double restriction enzyme digestion and DNA sequencing confirmed that the eukaryotic expression vector p DsRed-Monomer-N1-p24 was constructed successfully. Fluorescence microscope showed that p24-DsRed-Monomer was co-localized with TRIM22-EGFP in HEK293 T cells. Conclusion HIV capsid protein p24 is co-localized with TRIM22 in HEK293 T cells.