利用RACE方法克隆小麦β-1，3-葡聚糖酶基因的全长cDNA  

A full-length cDNA cloning of a wheat β-1,3-glucanase gene using rapid amplification of cDNA ends

作　　者：崔素萍[1] 刘博[1] 黄丽丽[1] 康振生[1] 

机构地区：[1]西北农林科技大学植物保护学院

出　　处：《西北农林科技大学学报（自然科学版）》2008年第7期79-84,共6页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家“863”项目“小麦抗病性状功能基因组研究”(2006AA10A104);国家“973”前期专项(2006CB708208);教育部重大科技培育项目(2004-295);教育部长江学者和创新团队发展计划项目(IRT0558);国家自然科学基金项目(30671350);高等学校学科创新引智计划项目(B07049)

摘　　要：【目的】研究小麦抗条锈菌的分子机制,在受条锈菌侵染的非亲和组合小麦叶片中克隆一个新的β-1,3-葡聚糖酶基因全长cDNA。【方法】采用RT-PCR、cDNA末端快速扩增技术(RACE),在感染小麦条锈菌的小麦(水源11)叶片中进行-β1,3-葡聚糖酶基因全长cDNA的扩增。【结果】得到了一个1338 bp-β1,3-葡聚糖酶基因全长cDNA,Genbank上注册号为DQ090946,测序结果和序列生物信息学分析结果表明,其与-β1,3-葡聚糖酶基因gi3757681和gi6782375的同源性分别为94.3%和92.1%。该全长cDNA具有完整的编码框,编码334个氨基酸,Genbank上注册号为AAY96422,与-β1,3-葡聚糖酶CAA77085和AAA32958的同源性分别为95.8%和86.7%。【结论】初步推断此全长cDNA是小麦中编码-β1,3-葡聚糖酶的一个新基因。[Objective] The paper studied the molecular mechanisms of wheat resistance to Puccinia striiformis. A full length cDNA of a β-1,3-glucanase gene was cloned in wheat leaves which were infected by Puccinia striiformis in incompatible interaction. [Method] RT-PCR and Rapid Amplification of cDNA Ends（RACE） technique was used to obtain a full length cDNA of a β-1,3-glucanase gene in wheat leaves which were infected by Puccinia striiformis. [Result] The full length cDNA has 1 338 bp nucleotides,the GenBank accession number is DQ090946,the sequence analysis result shows that it has 94.3% ,92.1% i- dentity with gi3757681, gi6782375, it has the complete open reading frame and is classified as 334 amino acids with codes, the GenBank accession number is AAY96422, it has 95. 8%, 86. 7% identity with CAA77085,AAM75342. [Conclusion] The full length cDNA is supposed to be a new gene of β-1,3-glu-canase Gene in Wheat leaves.

关 键 词：小麦 条锈菌 Β-1 3-葡聚糖酶基因 分子克隆 全长CDNA RACE 

分 类 号：S435.121.42[农业科学—农业昆虫与害虫防治]