RACE方法获得番茄蓝光受体基因Phototropin-2全长及其过量表达载体的构建  

Acquiring Full Length cDNA of Tomato Phototropin-2 with RACE PCR Technology and Construction of its Overexpression Vector

作　　者：喻堃[1] 牛向丽[1] 汪松虎[1] 

机构地区：[1]四川大学生命科学学院生物资源与生态环境教育部重点实验室,四川成都610064

出　　处：《安徽农业科学》2006年第22期5808-5809,5869,共3页Journal of Anhui Agricultural Sciences

摘　　要：采用RACE及降落PCR技术从番茄幼苗总RNA中克隆出PHOT-2基因的3’及5’末端片断,并根据测序结果,设计拉基因全长引物获得了PHOT-2基因全长。结果表明,该基因全长3 381 bp,含开放阅读框2 856 bp,编码952个氨基酸;与拟南芥PHOT-2氨基酸序列相比,同源性高达68%,主要功能区域同源性高达99%。将番茄PHOT-2基因定向插入pHB质粒中构建成过量表达载体,重组质粒转化农杆菌EHA105感受态细胞。Utilizing RACE and touchdown PCR technology, both 3＇and 5＇ cDNA ends of the blue light photoreceptor Pbotoropin-2 from tomato seedings were acquired. According to their sequence, the lull length cDNA of PHOT-2 was Cloned and sequenced. HHOT-2 cDNA fragment was 3 381 bp in length, including 2 856 bp of its ORF,encoding 952 amino acid. It shared 68% identity to Arabidopsis thaliana PHOT-2 in whole amino acid sequence and at the important action domain it ahnost shared 99% identity to Arabidopsis thaliana. Its overexpression vector was constructed by directionally inserting it into pHB plasmid and the recombinant plasmid was transformed into Agrobacterium tumefaciens EHA105 cells.

关 键 词：番茄 蓝光受体 RACE PCR技术 降落PCR技术 过量表达载体 

分 类 号：Q943.2[生物学—植物学]