层理鞭枝藻藻蓝蛋白β亚基的体内重组  

Reconstitution in vivo of C-Phycocyanin Polo-β Subunit from M.laminosus PCC 7603

作　　者：赵金梅[1] 朱菁萍[1] 王锋[1] 周明[1] 赵开弘[1] 

机构地区：[1]华中科技大学环境科学与工程学院,湖北武汉430074

出　　处：《武汉大学学报（理学版）》2006年第4期481-486,共6页Journal of Wuhan University:Natural Science Edition

基　　金：国家自然科学基金资助项目(30540070)

摘　　要：为研究藻蓝蛋白β亚基的生物合成途径,通过构建相容的3种重组质粒pET-cpcB(C155I)、pCDFDu-et-cpeS和pACYCDuet-ho1-pcyA,将裂合酶基因cpeS、血红素氧化酶基因ho1、藻蓝胆素合成酶基因pcyA和藻蓝蛋白β脱辅基蛋白基因cpcB(C155I)共同转入大肠杆菌.通过色素蛋白锌电泳和光谱检测,结果表明产生了有生物活性的CpcB(C155I)-PCB.而在裂合酶基因cpeS不转入大肠杆菌的情况下,大肠杆菌内只有7.6%的CpcB(C155I)-PCB产生.实现了大肠杆菌内藻蓝蛋白β亚基84位半胱氨酸残基与PCB连接,为进一步在大肠杆菌内合成天然的藻蓝蛋白奠定了基础.The entire pathway for the synthesis of a fluorescent holophycobiliprotein subunit from a photosynthetic cyanobacterium was reconstituted in E. coli. Cyanobacterial genes for enzymes hol and pcyA were expressed from a plasmid pACYCDuet-hol-pcyA. Genes for the apoprotein （C-phycocyaninβ subunit; cpcB（C155Ⅰ）） were expressed on a second plasmid pET-cpcB（C155Ⅰ）. Genes for the lyase （cpeS） that catalyzes chromophore attachment were expressed on a third plasmid pCDFDuet-cpeS. Upon induction,recombinant E. coli used the cellular pool of heme to produce CpcB（C155Ⅰ）-PCB with spectroscopic properties ,qualitatively and quantitatively similar to those of the same protein produced endogenously in cyanobacteria. About 7.56% of the CpcB（C155Ⅰ） was converted to CpcB（C155Ⅰ）-PCB in a similarly engineered E. coli strain that lacks cpeS.

关 键 词：C-藻蓝蛋白 藻蓝蛋白β脱辅基蛋白 裂合酶基因cpeS 体内重组 

分 类 号：Q93[生物学—微生物学] Q7