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.
出处
《安徽农业科学》
CAS
北大核心
2006年第22期5808-5809,5869,共3页
Journal of Anhui Agricultural Sciences
