东亚飞蝗血蓝蛋白亚基1基因的原核表达与蛋白结构预测  

Prokaryotic expression of the hemocyanin subunit type 1 gene from Locusta migratoria manilensis and protein prediction

作　　者：张新新[1] 游婷[1] 刘海霞[1] 印红[1] 

机构地区：[1]河北大学生命科学学院,河北保定071002

出　　处：《河北大学学报（自然科学版）》2013年第6期636-641,共6页Journal of Hebei University(Natural Science Edition)

基　　金：国家自然科学基金资助项目(31272293);河北省自然科学基金资助项目(C2013201246)

摘　　要：为了分析东亚飞蝗血蓝蛋白亚基1(Lmi Hc1)基因的原核表达情况,并对其蛋白结构进行预测,采用PCR方法扩增Lmi Hc1目的基因片段,克隆至pET-DsbA表达载体中,构建含目的基因的表达质粒pET-DsbA/Lmi Hc1,将重组体转化为E.coli BL21(DE3)并诱导蛋白表达,至此成功构建了Lmi Hc1的原核表达载体.SDS-PAGE电泳结果显示目的基因得到了相对分子质量约为60ku的高表达的融合蛋白.采用SABLE与Swiss-Model软件分别预测Lmi Hc1蛋白的二级结构和三级结构,Lmi Hc1蛋白的二级结构为α螺旋、β折叠、无规则卷曲的混合体;其三级结构含有3个结构域:Hemocyanin_N,Hemocyanin_M,Hemocyanin_C.The aim of this study was to investigate the prokaryotic expression of hemocyanin subunit type 1gene from Locusta migratoria manilensis and predict the protein structure.A recombinant strain which could express antigen of Lmi Hc1was constructed.Lmi Hc1gene was amplified by PCR.The DNA products of Lmi Hc1were inserted into a prokaryotic expression vector pET-DsbA,and then translated in- to E.coli strain BL21（DE3）.SDS-PAGE electrophoresis was analyzed.The prokaryotic expression vector of Lmi Hc1gene was successfully constructed.And the result of SDS-PAGE electrophoresis showed that the target was expressed highly and effectively.The relative molecular mass of product was approximately 60ku.The Lmi Hc1protein secondary and tertiary structure were separately predicted by using SABLE and Swiss-Model.Secondary structure was a mixture withα-helix,β-sheet and coil.Tertiary structure con- tained three structure domain：Hemocyanin_N,Hemocyanin_M,Hemocyanin_C.Lmi Hc1could be normal- ly expressed in E.coli and used for the further study of protein structure and function.

关 键 词：东亚飞蝗 血蓝蛋白亚基1 原核表达 蛋白结构预测 

分 类 号：Q786[生物学—分子生物学]