| 雄性育性基因RNA干扰载体的构建与鉴定 |
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| 引用本文： | 王文锋,王红霞,穆灵敏.雄性育性基因RNA干扰载体的构建与鉴定[J].农业科学与技术,2009,10(3):26-28. |
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| 作者姓名： | 王文锋 王红霞 穆灵敏 |
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| 作者单位： | 王文锋,王红霞,WANG Wen-feng,WANG Hong-xia(新乡医学院生命科学技术系,河南新乡,453003);穆灵敏,MU Ling-min(新乡医学院形态学实验室,河南新乡,453003) |
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| 摘 要： | 1材料与方法1．1材料酶和试剂：各种限制性内切酶购自Roche公司，T4DNA连接酶购自上海生工公司，Taq聚合酶、dNTP、凝胶回收试剂盒均购自Promega公司，引物由上海生工公司合成，其他试剂均为进口或国产分析纯。植物材料和质粒：棉花洞A雄性可育株由西南大学棉花教研室提供，表达载体P^BП21质粒和大肠杆菌菌株XL1-Blue由西南大学生物技术中心实验室保存。pUCm-T克隆载体购自上海生工公司。
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| 关 键 词： | 棉花 雄性育性基因 RNA 干扰载体 鉴定 |
Construction and Identification of Male Sterile RNAi Vector |
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| WANG Wen-feng,WANG Hong-xia,MU Ling-min.Construction and Identification of Male Sterile RNAi Vector[J].Agricultural Science ＆ Technology,2009,10(3):26-28. |
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| Authors: | WANG Wen-feng WANG Hong-xia MU Ling-min |
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| Institution: | 1, Department of Life Science and Technology, Xinxiang Medical College, Xinxiang 453003; 2, Laboratory of Morphology, Xinxiang Medica College, Xinxiang 453003) |
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| Abstract: | cDNA fragment of fertility gene MS2 from cotton was cloned by RT-PCR approach, it was highly homologous with relevant genes of Brassica napus and Arabidopsis thaliana. According to the principles of constructing RNAi vector, sense and antisense fragments of MS2 gene carrying restriction endonuclease recognition sites were amplified via PCR technique, ligated with the first intron of upland cotton chinase gene, then inserted into artificially modified plant expression vector pBI121, yielding RNAi vector pBGP12MSIn. The results showed that RNAi vector pBGP12MSIn harboring MS2 gene driven by anther specific promoter BGP was successfully constructed. Our results laid a foundation for studying the function of this gene and genetic transformation of plant male sterile lines. |
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| Keywords: | Cotton MS2 RT-PCR RNAi |
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