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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.)

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【作者】 康传红王淑春陈胜勇李彩凤

【Author】 KANG Chuan-Hong1,WANG Shu-Chun1,CHEN Sheng-Yong2,LI Cai-Feng2,1College of Life Sciences,Heilongjiang University,Harbin 150080,China; 2College of Agriculture,Northeast Agricultural University,Harbin 150030,China

【机构】 黑龙江大学生命科学学院东北农业大学农学院

【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.

【基金】 国家自然科学基金(30471017,30771276);黑龙江省博士后科研启动基金(BSH-Q06103)
  • 【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
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