种子特异性表达噬夏孢欧文氏菌crtB基因的植物表达载体构建  

Construction of Plant Expressing Vector Containing crtB Gene from Erwinia uredovora

作　　者：刘慧娟[1] 郎君[1] 冯志国[1] 何光源[1] 

机构地区：[1]华中科技大学生命科学与技术学院分子生物物理教育部重点实验室,武汉430074

出　　处：《生物技术通报》2009年第1期91-94,共4页Biotechnology Bulletin

基　　金：国家973计划项目(2002CB111300)

摘　　要：利用PCR技术扩增出tp和crtB基因,胶回收后分别连接到pMD18-T载体测序。tp和crtB基因经酶切回收后通过pBlue-scriptKS质粒连在一起构建成pSK-tp-crtB,通过BamHⅠ和SacⅠ酶切,从pSK-tp-crtB载体上切下目的基因片段tp-crtB,替换pBI121-napA载体中的GFP基因,构建了tp-crtB基因在植物种子中特异性表达的载体pBI121-tp-crtB。转化至大肠杆菌感受态细胞DH5α。测序鉴定后将阳性质粒转化农杆菌感受态细胞EHA105,经菌液和质粒PCR分析,获得了真核表达载体pBI121-tp-crtB,为crtB基因功能的进一步研究奠定基础。Both crtB and tp gene were amplified by PCR and cloned into pMD18-T vector respectively. The pMD18-tp and pMD18-crtB were digested and the crtB and tp gene were linked together by the pBlue-script KS vector. By digestion of pSK-tp-crtB and pBI121,the GFP gene in pBI121 was replaced with tp-ertB gene. The plant expression vector pBl121-tp-crtB was obtained,which was transformed into the E. coli component DH5α. The expressing vector was identified by sequencing and then transformed into Agrobacterium tum efaciens EHA105,which was constructed successfully by PCR identification. Therefore the construction of recombinant pBI121-tp-crtB plant expression vector provides a basis for the research on functions of crtB gene.

关 键 词：crtB基因 根癌农杆菌 遗传转化 植物表达载体 

分 类 号：Q943.2[生物学—植物学]