项 艳 ，张永风 ，江海洋 ，韩国民 ， 朱苏文 ，程备久*
（安徽农业大学 安徽 合肥 230036）
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pUCCRNAi载体上。利用 pHMW·GUS与pCAMBIA1300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pCAMBIA1300+HMW+2F。通过三亲杂交将pCAMBIA1300+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。
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Construction and Transformation of Maize Starch Branching Enzyme SBEⅡb Gene RNA Interference Expression Vector
XIANG Yan,， ZHANG Yong-Feng,， JIANG Hai-Yang,，
HAN Guo-Min ，ZHU Su-wen， CHENG Bei-jiu*
(Anhui Agricultural University , Hefei 230036，Anhui, China)
Abstract: Twentieth extron (155 bp) from genome of sbeⅡb 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.
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