| 稻瘟病新抗性基因Pi36候选基因双元载体的构建 |
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| 引用本文： | LIU Xin-qiong,刘新琼,王春台,刘学群,王玲. 稻瘟病新抗性基因Pi36候选基因双元载体的构建[J]. 湖北农业科学, 2007, 46(2): 169-173 |
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| 作者姓名： | LIU Xin-qiong 刘新琼 王春台 刘学群 王玲 |
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| 作者单位： | 中南民族大学生命科学学院生物技术国家民委重点实验室,武汉,430074;华南农业大学资源环境学院,广州,510642 |
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| 摘 要： | 为了验证Pi36候选基因的功能,利用长片段PCR(long-range PCR,LR-PCR)技术从Pi36基因的供体品种Q61中扩增了3个候选基因Pi36-1,Pi36-2,Pi36-3,长度分别为5.9 kb,9.6 kb和16.5 kb,电泳回收目的片段,将Pi36-1和Pi36-2分别克隆到双元载体pCAMBIA1300,而将Pi36-3克隆到双元载体pYLTAC27的AscI位点.为了将Pi36-3也克隆到具有高效转化效率的双元载体pCAMBIA1300中,首先在不改变pCAMBIA1300基本骨架的前提下,在其多克隆位点增加了AscI酶切位点,获得新载体pCAMBIA1300AscI;然后将克隆在载体pYLTAC27上的Pi36-3片段切下来,组装到新载体pCAMBI-A1300AscI上.经过酶切鉴定和测序分析,已成功地获得了3个候选基因的重组阳性克隆,为进一步地研究Pi36基因的功能奠定了基础.
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| 关 键 词： | 稻瘟病菌 候选抗性基因 长片段PCR技术 双元载体 |
| 文章编号： | 0439-8114(2007)02-0169-05 |
| 修稿时间： | 2007-01-15 |
Binary Vector Construction of Candidate Genes of the Blast Resistance Gene Pi36 |
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| LIU Xin-qiong. Binary Vector Construction of Candidate Genes of the Blast Resistance Gene Pi36[J]. Hubei Agricultural Sciences, 2007, 46(2): 169-173 |
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| Authors: | LIU Xin-qiong |
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| Affiliation: | 1. Key Biotechnology Laboratory of State Ethnic Affairs Commission, College of Life Science, South-central University for Nationalities, Wuhan,430073,China; 2. College of Resources and Environmental Sciences, South China Agricultural University,Guangzhou 510642,China |
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| Abstract: | To identify three candidate genes predicted,long-range PCR(LR-PCR) was employed to amplify each of the candidate genes based on the genomic DNA of the donor cv.Q61.The appropriate products of the Pi36-1 and Pi36-2 were purified and then ligated into the binary vector of pCAMBIA1300,as well as that of the Pi36-3 ligated into the binary vector of pLYTAC27 and the reconstructed pCAMBIA1300AscI,respectively.The results would aid to dissect the function of the resistance gene Pi36. |
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| Keywords: | Magnaporthe grisea candidate resistance gene long-range PCR binary vector |
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