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。
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.
出处
《植物生理学通讯》
CSCD
北大核心
2008年第4期710-714,共5页
Plant Physiology Communications
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)
关键词
甜菜
谷氨酰胺合成酶
基因克隆
sugar beet （Beta vulgaris）
glutamine synthetase
gene cloning
