摘要
利用 PCR技术从油菜 Polim a不育系中扩增出细胞质雄性不育相关基因 orf 2 2 4 ,并将其克隆到p GEM- T Easy载体上得到重组质粒 p GEMORF。在此基础上 ,用限制性内切酶将 orf 2 2 4基因从质粒 p GEMORF上切下 ,连接在 p BI12 1质粒的 Ca MV 35 S启动子和 NOS终止子之间。经 PCR和酶切鉴定 ,得到了 orf 2 2 4基因的植物表达载体 p
orf 224 was a CMS related gene discovered in Polima cytoplasm. In this paper,genomic DNA of Polima CMS line was used as templates,and two primers of specific oligonucleotides at 5′ and 3′end were used,PCR was performed. The amplification fragment was cloned into pGEM T Easy vectors and a recombinant plasmid pGEMORF with orf 224 was obtained.A plant expression vector was constructed by inserting orf 224 gene that cut from pGMORF using restriction endonuclease into the site between CaMV 35S promoter and NOS terminator of pBI121.PCR and restriction enzyme digestion analysis showed that the plant expression vector pBIORF containing orf 224 gene was obtained.This research created a new method and founded the basis for generating CMS line.
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2002年第4期9-12,共4页
Journal of Northwest A&F University(Natural Science Edition)
基金
高等学校博士学科点专项科研基金项目 (960 70 1)
