ULBP3胞外段分子伴侣10融合蛋白的构建与表达

Construction and expression of ULBP3 extracellular domain chaperonin10 recombinant fusion protein

[目的 ]研究ULBP3基因的功能 ,构建、表达ULBP3胞外段分子伴侣 10重组融合蛋白。 [方法 ]将ULBP3胞外段cDNA连接在重组原核表达质粒 pET2 8a -chaperonin10中chaperonin10基因的下游 ,构建chaperonin10 -ULBP3融合基因的 pET2 8a原核表达载体 ,转化大肠杆菌BL2 1(DE3) ,以IPTG诱导表达 ,Westernblot鉴定。 [结果 ]酶切鉴定证实 ,chaperonin10 -ULBP3融合基因的 pET2 8a原核表达载体构建正确 ,该原核表达载体转化大肠杆菌BL2 1(DE3)后 ,经筛选获得的阳性重组菌稳定表达chaperonin10 -ULBP3重组融合蛋白。 [结论 ]成功表达了ULBP3胞外段重组融合蛋白 ,可进一步用于ULBP3功能实验研究。

[Objective] To study the function of ULBP3 and express the ULBP3 extracellular domain recombinant fusion protein. [Methods] The extracellular domain of ULBP3 cDNA was cloned into the recombinant pET28a-chaperonin10 prokaryotic expression vector to construct a recombinant pET28a-chaperonin10-ULBP3 prokaryotic expression vector with ULBP3 cDNA fused to the hydroxy end of chaperonin10 gene. Then the recombinant plasmid was transduced into E.Coli. expression host BL21(DE3), the expression of fusion protein was induced by IPTG, and identified by Western blot. [Results] recombinant pET28a-chaperonin10-ULBP3 plasmid was constructed successfully and recombinant fusion protein chaperonin10-ULBP3 was identified by SDS-PAGE and Western blot. [Conclusion] The ULBP3 extracellular domain recombinant fusion protein was expressed in BL21(DE3) stably.