| 摘要: |
| 利用PCR技术从油菜Polima不育系中扩增出细胞质雄性不育相关基因orf224，并将其克隆到pGEM-T Easy载体上得到重组质粒pEGMORF。在此基础上，用限制性内切酶将orf224基因从质粒pGEMORF上切下，连接在pBI121质粒的CaMV 35S启动子和NOS终止子之间，经PCR和酶切鉴定，得到了orf224基因的植物表达载体pBIORF。 |
| 关键词: 细胞质雄性不育 orf224 基因克隆 植物表达载体 |
| DOI： |
| 分类号: |
| 基金项目:高等学校博士学科点专项科研基金项目(960701) |
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| Cloning of orf224 gene correlated with cytoplasmic male sterility (CMS)and its plant expression vector construction |
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| Abstract: |
| orf224 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 orf224 was obtained.A plant expression vector was constructed by inserting orf224 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 orf224 gene was obtained.This research created a new method and founded the basis for generating CMS line. |
| Key words: cytoplasmic male sterility orf224 gene cloning plant expression vector |
