发根农杆菌转化新疆紫草的PCR检测及序列分析  

Detection and Sequence Analysis on PCR of Arnebia euehroma Jonst Indueed by Agrobacterium rhizogenes

作　　者：芦韦华[1] 陈永芳[1] 代宁波[1] 李翠芳[1] 郝爱花[1] 王芳[1] 

机构地区：[1]新疆农业大学农学院,乌鲁木齐830052

出　　处：《新疆农业科学》2009年第1期63-66,共4页Xinjiang Agricultural Sciences

基　　金：国家自然科学基金项目(30560059);新疆农业大学青年教师科研启动基金项目

摘　　要：以发根农杆菌MSU440菌株转化的新疆紫草毛状根为材料,设计了rolC基因的特异性引物,应用PCR技术,扩增得到了600 bp左右的特异性核苷酸片段。进行双向测序分析,扩增的基因序列全长626 bp,含rolC基因的编码区540 bp,编码180个氨基酸组成的蛋白。正向测序rolC基因与GENEBENK已发表的序列相比较,核苷酸序列的同源性达到100%,氨基酸同源性高达100%;反向测序中,同源性达到97%。证实并确认了新疆紫草毛状根的真实性,为进一步利用新疆紫草毛状根生产次生代谢产物研究奠定了物质基础。Based on the analytical results of pRiA4 Ti - plasmid sequence in GENE - BANK, a fragment of about 600 bp was amplified from the Arnebia euchroma （Royle） Johust hairy roots were induced of Agrobacterium rhizogenes strain MSU440 by PCR method. Sequence Analysis of talC Gene was conducted. The results showed that the segment was 626 bp and contained one open reading frames that was composed of 540 bp nucleotides encoding a coat protein gene of 180 amino acids. Comparison with previously published nudeotide sequence showed that the homology was 100 % at nuleotide level, and 100 % at amino adde level; antisequence analysis compared with previously published nueleotide sequence showed that the homology was 97 %. From the above results, the Arnebia euchroma hairy roots were really induced by Agrobacterium rhizogenes .

关 键 词：新疆紫草 发根农杆菌 ROLC基因 序列分析 

分 类 号：S567.239[农业科学—中草药栽培]