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甜菜胞质型谷氨酰胺合成酶基因组DNA的克隆
Genomic DNA Cloning of Cytosolic Glutamine Synthetase from Sugar Beet (Beta vulgaris L.)
【摘要】 以甜菜叶片为材料,用CTAB法提取基因组DNA。以分段PCR法扩增得到了完整的甜菜胞质型谷氨酰胺合成酶(GS1)基因组DNA。采用RT-PCR法扩增此GS1基因(GS1)的cDNA序列应用于对照。获得了长度为9606bp的完整的GS1DNA序列和长度为1068bp的GS1cDNA序列。分析GS1基因组DNA序列表明,它包含13个外显子,被12个内含子分隔开。其外显子区与已公布的GS1mRNA序列的相似性达99.5%。RT-PCR法获得的cDNA序列与已知的GS1mRNA序列相似性达99.6%。而2次实验中GS1基因组DNA外显子区与GS1cDNA序列的相似性达99.9%。GenBank登录号为EU370974。
【Abstract】 Genomic DNA was extracted from sugar beet leaves by CTAB method in this paper. Cytosolic glutamine synthetase (GS1) genomic DNA was cloned by subsection PCR method,and the full-length GS1 gene sequence was successfully obtained. The partial GS1 cDNA sequence was obtained by RT-PCR method and compared to GS1 gene. The full length of GS1 genomic DNA was 9 606 bp,including 13 exons which were separated by 12 introns. The sequence similarity between exon parts of GS1 genomic DNA and the known GS1 mRNA was 99.5%. GS1 cDNA sequence was 1 068 bp,and the sequence similarity between this GS1 cDNA and the known GS1 mRNA was 99.6%. Meanwhile,the sequence similarity between exon parts of GS1 genomic DNA and GS1 cDNA cloned in the experiment was 99.9%. The gene accession number in GenBank was EU370974.
【Key words】 sugar beet (Beta vulgaris); glutamine synthetase; gene cloning;
- 【文献出处】 植物生理学通讯 ,Plant Physiology Communications , 编辑部邮箱 ,2008年04期
- 【分类号】Q943.2;S566.3
- 【被引频次】3
- 【下载频次】189