块根特异启动的木薯SBE Ⅰ、SBE Ⅱ基因RNAi载体的构建及鉴定  

Construction and Identification of RNAi Vector Containing Cassava SBE Ⅰ,SBE Ⅱ Gene and Root Specific Promotor

作　　者：弓淑芳[1,2] 郭安平[2] 郭运玲[2] 孔华[2] 阳辛凤[2] 贺立卡[2] 

机构地区：[1]海南大学农学院,海南海口570228 [2]中国热带农业科学院热带生物技术研究所,海南海口571101

出　　处：《热带作物学报》2011年第8期1507-1511,共5页Chinese Journal of Tropical Crops

基　　金：国家自然科学基金项目(No.30960204);国家重点基础研究发展计划(973)项目(No.2010CB126605);中央级公益性科研院所基本科研业务费(No.ITBB110601)

摘　　要：利用基因重组技术,分别克隆木薯SBEⅠ基因ORF中的494 bp的片段和SBEⅡ基因ORF中的340 bp的片段;并通过重叠延伸PCR方法,扩增融合SBEⅠ、SBEⅡ基因片段的大片段SⅢ,将其正向、反向插入植物RNAi表达载体pART27中,同时克隆块根特异性启动子取代原来载体上自有的35S启动子,构建块根特异启动的可编码发夹结构的RNAi表达载体Sp-pRNAiSⅢ,为后续培育高直链淀粉含量的木薯新品种等实验打下基础。A 494 bp fragment named SI from the Open Reading Frame（ORF）of SBE I and a 340 bp fragment named S II from the ORF of SBE II were cloned from cassava and fused into a larger fragment named SIII with Overlap Extension PCR（SOE PCR）. The SⅢ fragment was inserted into the plant expression vector pART27 RNAi in forward and reverse direction; Also the original 35S promotor in the vector was replaced with a root specific promotor Sporamin, and then a coding hairpin RNAi expression vector Sp-pRNAiS Ⅲ was constructed, which could, lay a foundation to breed new cassava varieties with high amylase content.

关 键 词：木薯 高直链淀粉 RNA干扰技术 重叠延伸PCR 

分 类 号：S533[农业科学—作物学] Q78[生物学—分子生物学]