玉米淀粉分支酶基因SBEⅡb的克隆与过表达载体的构建

中国农学通报 ›› 2008, Vol. 24 ›› Issue (4): 72-75.

所属专题：生物技术；玉米

玉米淀粉分支酶基因SBEⅡb的克隆与过表达载体的构建

范亚丽, ,阮颖,李进, ,杜培粉, ,姚远颋, ,刘春林

收稿日期:2008-12-07 修回日期:2008-01-15 出版日期:2008-04-05 发布日期:2008-04-05

Cloning of the Maize Starch Branching Enzyme SBEⅡb Gene and Constructing of its Over-expression Vector

Fan Yali ,, Ruan Ying ,Li Jin ., Du Peifen, , Yao Yuanting, , Liu Chunlin

Received:2008-12-07 Revised:2008-01-15 Online:2008-04-05 Published:2008-04-05

摘要/Abstract

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（Waxy corn）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。

关键词: 室内观赏植物, 室内观赏植物, 甲醛, 吸纳

Abstract: 【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.

范亚丽, ,阮颖,李进, ,杜培粉, ,姚远颋, ,刘春林. 玉米淀粉分支酶基因SBEⅡb的克隆与过表达载体的构建

[J]. 中国农学通报, 2008, 24(4): 72-75.

Fan Yali ,, Ruan Ying ,Li Jin ., Du Peifen, , Yao Yuanting, , Liu Chunlin. Cloning of the Maize Starch Branching Enzyme SBEⅡb Gene and Constructing of its Over-expression Vector[J]. Chinese Agricultural Science Bulletin, 2008, 24(4): 72-75.

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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

Cloning of the Maize Starch Branching Enzyme SBEⅡb Gene and Constructing of its Over-expression Vector

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