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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
【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。
【Abstract】 A 494 bp fragment named SI from the Open Reading Frame(ORF)of SBEⅠ and a 340 bp fragment named SⅡ from the ORF of SBEⅡwere 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-pRNAiSⅢ was constructed,which could,lay a foundation to breed new cassava varieties with high amylase content.
【Key words】 Cassava(Manihot esculenta Crantz); high-amylose starch; RNAi; splicing by Overlap Extension PCR;
- 【文献出处】 热带作物学报 ,Chinese Journal of Tropical Crops , 编辑部邮箱 ,2011年08期
- 【分类号】S533
- 【被引频次】4
- 【下载频次】121