在链霉菌中表达透明颤菌血红蛋白需要异源启动子  

Heterologous Promoter is Necessary for the Expression of Vitreoscilla Hemoglobin in Streptomyces

作　　者：崔风文[1] 杨胜利[1] 

机构地区：[1]中国科学院上海生物工程研究中心

出　　处：《生物工程学报》1998年第1期91-95,共5页Chinese Journal of Biotechnology

摘　　要：构建了质粒ｐＩＪ４０８３Ｍｐｒｏ、ｐＩＪ４０８３ｐｒｏ、ｐＷＬＤ８和ｐＦＷ３。在浅青紫链霉菌ＴＫ２４中，启动子探针质粒ｐＩＪ４０８３上的邻苯二酚双加氧酶基因（ｘｙｌＥ）不能被透明颤菌血红蛋白基因（ｖｇｂ）的启动子带动转录，表明ｖｇｂ启动子在链霉菌中无作用。ＴＫ２４中，ｐＷＬＤ８和ｐＦＷ３均能表达透明颤菌血红蛋白（ＶＨｂ），ｐＷＬＤ８上可能是由Ｐｌａｃ带动ｖｇｂ的表达；ｐＦＷ３上ｖｇｂ基因去掉了非必要部分，克隆在ＰＣＲ扩增得到的ｇｌｎＡ启动子下游，两者连成嵌合基因。Plasmids pIJ4083Mpro,pIJ4083 pro,pWLD8 and pFW3 were constructed to study Vitreoscilla hemoglobin(VHb) expression in Streptomyces .The xylE gene on pIJ4083 could not be transcribed by Vitreoscilla hemoglobin gene(vgb) promoter in Streptomyces lividans TK24,it indicated that vgb promoter was functionally inactive in Streptomyces. pWLD8 and pFW3 expressed VHb in TK24.Probably Plac is responsible for VHb expression on pWLD8,while on FW3 vgb gene deleted of its unnecessary parts were cloned down stream from glutamine synthetase gene(glnA)promoter amplified through PCR to form a chimerical gene with PglnA.

关 键 词：浅青紫链霉菌 透明颤菌血红蛋白 异源表达 供氧 

分 类 号：TQ465[化学工程—制药化工]