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玉米淀粉分支酶基因SBEⅡb的克隆与过表达载体的构建 被引量:3

Cloning of the Maize Starch Branching Enzyme SBEⅡb Gene and Constructing of its Over-expression Vector
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摘要 【研究目的】克隆玉米分支酶基因SBEⅡb并构建该基因的过表达载体pCASBEⅡb。【研究方法】从糯玉米(Waxycorn)新鲜胚乳中提取总RNA,利用RT-PCR方法克隆玉米淀粉分支酶基因SBEⅡb的全长cDNA。【结果】克隆得到了玉米淀粉分支酶基因SBEⅡb的全长cDNA,用BLAST软件作同源序列比对的结果显示,该片段的核苷酸序列与GenBank上报道的序列具有99%的同源性,全长2719bp,编码799个氨基酸。【结论】将该基因连接到携带GUS报告基因的植物表达载体pCAMBIA1303中,并通过了GUS活性检测,说明已成功构建了玉米淀粉分支酶基因SBEⅡb的过表达载体pCASBEⅡb。 [OBJECTIVE]In this article we cloned the Maize Starch Branching Enzyme SBEⅡb Gene and constructed its over-expression vector successfully. [METHOD]The total RNA extracted from Waxy corn endosperm was as template, the Coding Regions of cDNA of the Maize Starch Branching Enzyme SBEⅡb gene was amplified Utilizing the method of RT-PCR.[RESULTS]The gene was successfully amplified. Homologous Analysis result by BLAST showed that the cloned fragment has 99% identity to the sequence of the gene in GenBank. The Coding Regions of SBEⅡb gene cDNA was 2719bp in length and encode 799 amino acids. [CONCLUSION]The cloned cDNA was ligated into the plant express-vector pCAMBIA1303 . We successfully constructed a gene over-expression vector pCASBEⅡb through GUS activity test.
出处 《中国农学通报》 CSCD 2008年第4期72-75,共4页 Chinese Agricultural Science Bulletin
基金 湖南省科技计划重点项目"马铃薯种质资源创新研究"(03NK1001)
关键词 玉米胚乳 淀粉分支酶基因 过表达载体 maize endosperm, Starch Branching Enzyme gene, over-express vector
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