G418抗性基因nptⅡ双元载体的构建及谢瓦氏曲霉间型变种的遗传转化

为给谢瓦氏曲霉间型变种（Aspergillus chevalieri var. intermedius）多基因功能的研究提供新的选择标记，利用MYA培养基测G418敏感性后，以质粒pcDNA3．1（＋）为中间载体构建倒Ⅱ基因表达元件，利用根癌农杆菌介导法转化至谢瓦氏曲霉间型变种。结果表明：用构巢曲霉PgpdA启动子、nptⅡ基因和构巢曲霉色氨酸c终止子TtrpC组成的G418抗性基因盒替换双元载体pDHt／sk-bar上的草丁膦抗性盒，成功构建双元载体pDHt／sknt，并且通过根癌农杆菌介导的转化获得的G418抗性转化子遗传稳定，nptⅡ基因主要以单拷贝形式插入谢瓦氏曲霉间型变种基因组DNA...

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Bibliographic Details
Published in西南农业学报 Vol. 26; no. 6; pp. 2509 - 2513
Main Author 曹旸 刘永翔 谭玉梅 王海 刘作易
Format Journal Article
LanguageChinese
Published 贵州省农业生物技术重点实验室,贵州贵阳550006 2013
贵州省农业生物技术重点实验室,贵州贵阳550006%贵州省农业生物技术重点实验室,贵州贵阳,550006%华中农业大学生命科学技术学院,湖北武汉430070
华中农业大学生命科学技术学院,湖北武汉,430070%贵州省农科院生物技术研究所,贵州贵阳550006
Subjects
C418抗性
双元载体
谢瓦氏曲霉间型变种
遗传转化
G418 resistance
Aspergillus chevalieri var.intermedius
Genetic transformation
遗传转化
Binary vector
双元载体
谢瓦氏曲霉间型变种
G418抗性
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ISSN1001-4829

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Summary:为给谢瓦氏曲霉间型变种（Aspergillus chevalieri var. intermedius）多基因功能的研究提供新的选择标记，利用MYA培养基测G418敏感性后，以质粒pcDNA3．1（＋）为中间载体构建倒Ⅱ基因表达元件，利用根癌农杆菌介导法转化至谢瓦氏曲霉间型变种。结果表明：用构巢曲霉PgpdA启动子、nptⅡ基因和构巢曲霉色氨酸c终止子TtrpC组成的G418抗性基因盒替换双元载体pDHt／sk-bar上的草丁膦抗性盒，成功构建双元载体pDHt／sknt，并且通过根癌农杆菌介导的转化获得的G418抗性转化子遗传稳定，nptⅡ基因主要以单拷贝形式插入谢瓦氏曲霉间型变种基因组DNA中。因此，双元载体pDHt／sknt适用于谢瓦氏曲霉间型变种的遗传转化。
Bibliography:G418 resistance; Binary vector;Aspergillus chevalieri vat. intermedius ; Genetic transformation
51-1213/S
CAO Yang, LIU Yong-xiang, TAN Yu-mci, WANG Hai, LIU Zuo-yi （1. College of Life Science and Technology, Huazhong Agricultural University, Hubei Wuhan 430010,China;2. Guizhou Institute of Bioteehnology,Guizhou Guiyang 550006 ,China;3. Guizhou Key Laboratory of Agricultural Bioteehnology, Guizhou Guiyang 55006, China）
In order to find a new selection marker for the study of multl-genes in A. chevallerl var. intermedius,the authors developed a npt Ⅱ gene expression cassette in plasmid pcDNA3. 1 （ ＋ ） after testing the sensitivity of G418, and utilizing the method of Agrobacterium tumefac/ens mediated transformation. The results showed that the binary vector pDHt/sknt was constructed in binary vector pDHt/sk-har while the nptII gene expression cassette,which was made up of promotor PgpdA, gene npt Ⅱ and terminator TtrpC, replaced the bar gene expression cassette, and it was proved that G418 resistance transfroman
ISSN:1001-4829