玉米淀粉分支酶SBEⅡb基因RNA干涉表达载体的构建和转化  

Construction and Transformation of Maize Starch Branching Enzyme SBEⅡb Gene RNA Interference Expression Vector

作　　者：项艳[1] 张永凤[1] 江海洋[1] 韩国民[1] 朱苏文[1] 程备久[1] 

机构地区：[1]安徽农业大学林学与园林学院,安徽合肥230036

出　　处：《激光生物学报》2006年第3期288-293,共6页Acta Laser Biology Sinica

基　　金：国家转基因植物研究与产业化专项基金(JY04-A-01)

摘　　要：应用聚合酶链式方应技术扩增玉米淀粉分支酶第20外显子155 bp的基因片段,并将其正向和反向克隆到pUCCRNA i载体上。利用pHMW.GUS与pCAMB IA1300作为桥梁载体,构建了含胚乳特异启动子和潮霉素筛选基因的RNA干涉表达载体pCAMB IA1300+HMW+2F。通过三亲杂交将pCAMB IA1300+HMW+2F表达载体转化到根癌农杆菌LBA4404,对玉米自交系178进行遗传转化,通过抗性筛选和PCR检测,获得了4株PCR阳性苗。Twentieth extron （155 bp） from genome of sbe H b gene have been amplified. The positive oriention and negative oriention of the fragment is ligated into pUCRNAi. Vector p1300 ＋ HMW ＋ 2F have been constructed by utilizing pHMW · GUS and pCAMBIA1300 vector which respectively contain hygromycin selecting gene. Through the tri-parental mating experiment, the RNAi expression vector pCAMBIA1300 ＋ HMW ＋ 2F has been transformed into A. tumefaciens LBA4404. Genetic transformation into maize inbred lines 178, four transgenic maize are obtained by hygromycin selecting and PCR analysis.

关 键 词：淀粉分支酶基因 小干涉RNA 表达载体的构建 玉米 农杆菌介导 

分 类 号：Q78[生物学—分子生物学] S513[农业科学—作物学]