摘要
采用PCR技术,从马铃薯叶片基因组DNA中扩增到了约0.6kb的马铃薯GBSS基因的5'侧翼序列,经序列同源性分析所克隆的序列与已知序列同源性为97.41%,含有启动基因表达所需的主要调控元件CAAT盒和TATA盒。构建该序列与GFP的融合基因植物表达载体pBIGGFP,并用基因枪转化法将pBIGGFP导入马铃薯无菌苗茎段、叶片和微型薯薯片,用荧光显微镜观察转化组织中GFP的瞬时表达,结果表明GFP在上述器官中都得到了表达,说明所克隆的约0.6kbGBSS基因的5'侧翼序列是具有启动基因表达的功能。
In this study, a 0.6 kb 5'flanking regions of GBSS gene was cloned from potato (Solanum Tuberosum L.) leaf genomic DNA via polymerase chain reaction (PCR). Sequence analysis indicates that the cloned sequence shares a 97.41% homology with the reported sequence (gi:1247300), and that it contains two main regulation domains, TATA box and CAAT box. However, there are a 14 bp insertion and a 136 bp deletion when compared with the reported sequence. In order to assay its regulation function, a plant expression vector pBIGGFP constructed by fusing the cloned sequence with GFP is transferred into stem segments, leaf discs and in viro microtuber slices via the particle bombardment. The transient expression of GFP proves that the cloned sequcence is functional.
出处
《中国马铃薯》
2005年第3期129-133,共5页
Chinese Potato Journal
基金
国家自然科学基金项目(项目批准号:30471101)
关键词
马铃薯
GBSS基因5’侧翼区
克隆
GFP
调控
potato (Solanum Tuberosum L.)
5' flanking sequence of granule-bound starch synthase gene
cloning
green flurescence protein
regulation
