HTLV-I gp21外膜蛋白基因片段的克隆和融合表达

Cloning and Fusion Expression of the gene encoding part of HTLV envelope protein gp21

目的 克隆HTLV 1 gp2 1外膜蛋白基因片段并在 pGEX 2T载体中融合表达。 方法 人工合成 4条寡核苷酸片段 ,串联成HTLV Igp2 1外膜蛋白基因片段 ,然后克隆入T载体 ,进行序列分析 ;再将序列正确的HTLV Igp2 1外膜蛋白基因片段亚克隆入pGEX 2T融合表达载体中 ,进行融合表达 ,并对表达的融合蛋白进行鉴定。 结果 序列分析结果显示 :利用人工合成的寡核苷酸片段串联成HTLV Igp2 1外膜蛋白基因片段是可行的 ;HTLV Igp2 1外膜蛋白基因片段亚克隆入pGEX 2T融合表达载体中得到了融合表达 ,且具有抗原性。 结论 具有特异抗原性的HTLV Igp2 1融合蛋白的成功表达为HTLV诊断试剂盒的研制奠定了基础 ,对保障输血安全具有意义。

Objective The gene encoding part of HTLV(Human Tcell leukemia viruses) envelope protein gp21 was cloned and expressed in order to prepare diagnostic agent for detecting HTLV.Methods The gene encoding part of HTLV envelope protein gp21 was obtained by combining four oligonucleotides into a long DNA fragment,then cloned and subcloned into Tvector and pGEX2T respectively.Expression was performed under induction in a fusion way that target protein gene was inserted downstream the carrying protein gene of pGEX2T which makes purification of target protein convenient.At last,Target protein was purified by affinity chromatography in glutathione sepharose 4B column and its antigenicity was confirmed by immunoblotting.Results and conclusion Sequence analysis showed it was feasible to obtain a target gene by ligating short oligonucleotides;As expected,target protein was expressed which exhibited strong antigenicity after being subcloned into fusion expression vector pGEX2T.