| OsWRKY10基因特异性dsRNA干涉载体构建 |
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| 引用本文： | 窦彩虹,陈应武.OsWRKY10基因特异性dsRNA干涉载体构建[J].安徽农业科学,2008,36(7):2682-2683. |
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| 作者姓名： | 窦彩虹 陈应武 |
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| 作者单位： | 陈应武,河南科技大学林学院,河南洛阳,471003;陈应武,河南科技大学林学院,河南洛阳,471003 |
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| 基金项目： | 河南科技大学人才科学研究基金，国家自然科学基金项目(30370139、30471122) |
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| 摘 要： | 目的]明确Osdg中T-DNA插入位点的基因是否为引起突变表型的基因。方法]采用OsWRKY10基因5’端、不含WRKY保守域的长300 bp的cDNA片段,构建抑制OsWRKY10基因表达的特异性dsRNA干涉重组表达载体pCam-35SWInW。采用冻溶法将pCam-35SWInW载体导入根癌农杆菌超毒力菌株EHA105,对其进行PCR检测。结果]该研究成功构建了重组表达载体pCam-35SWInW。采用相应限制性内切酶对重组表达载体进行酶切鉴定,其中以XhoⅠ消解重组质粒,可获得1 kb左右的DNA片段。在抑制基因表达方面,发卡结构比反义结构更为有效。结论]该研究成功构建了抑制OsWRKY10表达的特异性dsRNA干涉载体,可用于进一步的基因功能研究。
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| 关 键 词： | OsWRKY10基因 特异性dsRNA干涉载体 构建 |
| 文章编号： | 0517-6611(2008)07-02682-02 |
| 修稿时间： | 2008年2月18日 |
Construction on Specific dsRNA Interference Vector of OsWRKY10 Gene |
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| DOU Cai-hong.Construction on Specific dsRNA Interference Vector of OsWRKY10 Gene[J].Journal of Anhui Agricultural Sciences,2008,36(7):2682-2683. |
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| Authors: | DOU Cai-hong |
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| Abstract: | Objective] The research aimed to definitude whether the T-DNA insert locus gene in Osdg was the gene that caused mutant phenotype.Method] cDNA fragment with the length of 300 bp on 5' end of OsWRKY10 gene without containing conserved domain WRKY was used to construct the specific dsRNA interference recombinant expression vector pCam-35SWInW that could inhibit the expression of OsWRKY10 gene.pCam-35SWInW vector was introduced into supervirulent strain EHA105 of Agrobacterium tumefaciens by freeze thaw method and it was detected by PCR.Result] The research successfully constructed the recombinant expression vector pCam-35SWInW.The enzyme digestion of the recombinant expression vector was made by using the corresponding restriction endonuclease for identifying,in which the recombinant plasmid was digested by XhoⅠ and DNA fragment with the length of about 1 kb was obtained.For inhibiting the gene expression,hairpin structure was more effective than antisense structure.Conclusion] The research successfully constructed the specific dsRNA interference vector that could inhibit the expression of OsWRKY10,which could be used in the further research of gene function. |
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| Keywords: | OsWRKY10 gene Specific dsRNA interference vector Construction |
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