甘薯ADP-葡萄糖焦磷酸化酶两个α亚基基因的克隆分析和植物表达载体的构建  

Cloning of Two α-Subunit Genes of ADP-Glucose Pyrophosphorylase from Sweet Potato and Plant Expression Cassettes Construction

作　　者：张聪[1,2] 郑雪莲[3] 蒲志刚[1] 吴洁[1] 王大一[4] 阎文昭[1] 

机构地区：[1]四川省农业科学院生物技术核技术研究所,四川成都610066 [2]西南大学园艺园林学院,重庆北碚400715 [3]电子科技大学生命科学与技术学院,四川成都610054 [4]四川省农业科学院作物研究所,四川成都610066

出　　处：《西南农业学报》2010年第3期619-624,共6页Southwest China Journal of Agricultural Sciences

基　　金：国家科技支撑计划(2007BAD78B00);国家863项目(2006AA100107);四川省财政育种工程专项资助

摘　　要：ADP-葡萄糖焦磷酸化酶(AGPase)催化ADP-葡萄糖合成反应,是淀粉生物合成的限速酶之一。从高淀粉甘薯品种川薯34总RNA逆转录的cDNA中克隆了AGPa1和AGPa2两个α亚基编码序列,进行生物信息学分析后,插入植物高效表达载体pCamb ia1301构建了pC-AGPa1和pC-AGPa2两个双元表达载体,并导入根癌农杆菌EHA105中。获得的工程菌菌株可用于遗传转化甘薯、马铃薯和木薯等重要的薯类作物,为薯类作物高淀粉育种奠定了基础。ADP-Glucose pyrophosphorylase（AGPase） was the enzyme responsible for the production of adenosine 5＇-diphosphase glucose（ADPGlc）.The AGPase reaction was the first committed step in the biosynthesis of starch,and AGPase was one of the key enzymes in this pathway.Total RNA was purified from high-starch sweet potato variety Chuanshu 34,and was reverse-transcripted into cDNAs.Two α-subunit genes of AGPase,AGPa1 and AGPa2,were amplified from cDNAs.Afterwards AGPa1 and AGPa2 were sub-cloned into pCambia1301 to construct the recombinant vectors pC-AGPa1 and pC-AGPa2 after bioinformatic analysis.pC-AGPa1 and pC-AGPa2 vectors were introduced into A.tumefaciens strain EHA105,and ready to be used in transformation of sweet potato,potato and cassava.

关 键 词：甘薯[Ipomoea batatas(L.) Lam.] ADP-葡萄糖焦磷酸化酶 淀粉合成 载体构建 

分 类 号：S531[农业科学—作物学]