BPI_(23)-Fcγ1重组融合蛋白在E.coli中的表达  

Expression of recombinant BPI_(23)-Fcγ1 fusion protein in E. coli

作　　者：柯岩[1] 安云庆[1] 杨贵贞[2] 

机构地区：[1]首都医科大学微生物学和免疫学教研室 [2]白求恩医科大学免疫学教研室

出　　处：《中华微生物学和免疫学杂志》2000年第5期451-455,共5页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金!(395 70 6 6 6 )

摘　　要：目的　构建pBV BPI60 0 Fcγ170 0 重组表达载体 ,转化E .coliDH5α诱导表达BPI2 3 Fcγ1抗菌重组蛋白。方法　采用RT PCR技术 ,从HL 6 0细胞和正常人白细胞mRNA中扩增编码BPI2 3 和IgG1Fc(Fcγ1)的基因 ;通过连接反应构建重组克隆载体和重组表达载体 ;转化感受态E .coliDH5α细胞 ,通过温控诱导表达BPI2 3 Fcγ1重组蛋白。结果　 (1)RT PCR获得预期的扩增产物BPI60 0bp和Fcγ170 0bp基因片段 ;(2 )成功构建pUC18 BPI180 、pUC18 BPI4 2 0 、pUC18 Fcγ170 0 重组克隆载体 ,DNA测序结果与文献报道一致 ;(3)成功构建pBV BPI60 0 Fcγ170 0 重组表达载体 ,酶切图谱分析与预期结果相符 ;(4 )转化感受态E .coliDH5α细胞 ,通过温控诱导表达BPI2 3 Fcγ1重组蛋白 ,表达量约占菌体蛋白总量的 2 0 %～ 30 % ;(5 )复性后重组蛋白具有抗菌活性和结合蛋白A的作用。结论　pBV BPI60 0 Fcγ170 0 重组表达载体构建成功 ;在E .coli中得到表达 ,获得具有抗菌活性的BPI2 3Objective To express BPI 23 Fcγ1 anti bacteria recombinant fusion protein. Methods BPI 23 and Fcγ1 genes were isolated by RT PCR from HL 60 and normal human leukocytes and inserted into expression vector pBV and the fusion protein expressed in E. coli DH5α in a temperature induced manner. Results The expressed BPI 23 Fcγ1 fusion protein constituted 20%～30% of total bacterial protein. The renatured BPI 23 Fcγ1 recombinant protein showed biological activities of anti bacteria and was able to form conjugate with protein A. Conclusion pBV BPI 600 Fcγ1 700 recombinant expression vector was successfully constructed, and BPI 23 Fcγ1 recombinant protein with anti bacteria activities was expressed in E. coli .

关 键 词：BPI23-Fcγ1重组蛋白 聚合酶链反应 抗菌蛋白 

分 类 号：R392[医药卫生—免疫学]