HIV-1 CRF07_BC gp41重组抗原在不同体系表达效率的比较

Comparison of expression efficiency of HIV-1 CRF07_BC gp41 recombinant antigen in different expression system

目的获得HIV-1 CRF07_BC株gp41重组抗原表达效率最佳的体系。方法采用基因工程技术将gp41重组蛋白基因分别构建到pThioHis A,pThioHis A-1(不含标签),PBV220,PGEX-6p-1原核表达载体。转化大肠杆菌BL21和Rosetta两种宿主菌,通过IPTG诱导表达并经WB鉴定。结果 4种原核表达质粒构建完成,蛋白成功表达。PGEX-6p-1表达量明显高于pThioHis A载体;而无标签载体组中,PBV220表达量优于pThioHis A-1。BL21和Rosetta宿主菌对表达体系无影响。结论 HIV-1 CRF07_BC株gp41重组抗原在大肠杆菌中以包涵体形式存在,构建在PGEX-6p-1和PBV220载体上gp41重组蛋白表达效率较高,为gp41疫苗的研究和血清学检测提供充足的抗原。

Objective To obtain the HIV- 1 CRF07_BC gp41 recombinant antigen plasmid with high expressionefficiency.Methods The expression vector including pThioHis A with tag and without tag, PBV220 and PGEX- 6p- 1encoding the gp41 protein were constructed. After transformed into the host cell like BL21 or Rosetta, the production of gp41protein was induced by adding IPTG and identified by Western- blot. Results Four kinds of vector were constructedcorrectly and the protein were expressed. With the fusion protein tag, the PGEX-6p-1 vector can produce more protein asinclusion bodies than pThioHis A;without the fusion protein tag, the PBV220 was better than the altered pThioHis A vector.The host cell did not influence the protein expression efficiency. Conclusion The HIV-1 CRF07_BC gp41 recombinantantigen is produced in host cell as inclusion bodies and the PGEX- 6p- 1 vector or the PBV220 is adapted for proteinexpression either with the fusion protein tag or without,these gp41 proteins could be an important source of antigens for vaccineresearch and diagnostic reagents development.