| Hs1 pro-1双子叶表达载体构建和转化大豆研究初报 |
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| 引用本文： | 陈平华,陈如凯,潘大仁,许莉萍,袁照年.Hs1 pro-1双子叶表达载体构建和转化大豆研究初报[J].福建农林大学学报(自然科学版),2003,32(4):486-489. |
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| 作者姓名： | 陈平华 陈如凯 潘大仁 许莉萍 袁照年 |
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| 作者单位： | 福建农林大学农业部甘蔗生理生态与遗传改良重点开放实验室,福建,福州,350002 |
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| 基金项目： | 国家863计划(2001AA241191,2002AA241031)，国家自然科学基金(39870545)资助项目. |
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| 摘 要： | 用从德国引进的抗线虫基因Hs1pro-1构建表达载体并转化大豆,以获得抗线虫转基因植株.提取克隆载体p1832,用Nco 酶切、Klenow补平和Sac 酶切,回收基因片段;提取表达载体pBI121,用Sma 和Sac 酶切,回收大片段;目的片段用T4DNA连接酶连接并转化E.coli,鉴定重组质粒;用基因枪轰击转化大豆品种早熟1号,获得18株再生苗,其中3株经PCR检测呈阳性,通过Southern杂交,证明Hs1pro-1基因已整合到其中2株大豆的基因组中.
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| 关 键 词： | Hs1pro-1 表达载体 构建 基因枪轰击 大豆 |
| 文章编号： | 1006-7817(2003)04-0486-04 |
| 修稿时间： | 2003年7月1日 |
Construction of Hs1 pro-1 dicotyledonous expression vector and transformation of soybean |
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| CHEN Ping-hua,CHEN Ru-kai,PAN Da-ren,XU Li-ping,YUAN Zhao-nianry of Agriculture,P.R.China,Fujian Agriculture and Forestry University,Fuzhou,Fujian ,China.Construction of Hs1 pro-1 dicotyledonous expression vector and transformation of soybean[J].Journal of Fujian Agricultural and Forestry University,2003,32(4):486-489. |
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| Authors: | CHEN Ping-hua CHEN Ru-kai PAN Da-ren XU Li-ping YUAN Zhao-nianry of Agriculture PRChina Fujian Agriculture and Forestry University Fuzhou Fujian China |
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| Institution: | CHEN Ping-hua,CHEN Ru-kai,PAN Da-ren,XU Li-ping,YUAN Zhao-nianry of Agriculture,P.R.China,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China) |
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| Abstract: | The expression vector of Hs1 pro-1 cloned by German scientists was constructed and transformed into soybean to get transgenic plants. Cloning vector p1832 was digested with restriction nuclease NcoⅠ, DNA polymerase Ⅰ Large Fragment and SacⅠ in turn. Expression vector pBI121 was digested with SmaⅠ and SacⅠ. The short fragment of p1832 and the large fragment of pBI121 were isolated respectively. The two fragments were ligated by T_4 DNA ligase and the reactant was transferred into E.coli. The recombinant plasmid was identified and transformed into soybean genotype Zaoshu 1 via particle bombardment. Eighteen kanamycin-resistant soybean seedlings were obtained and three seedlings were positive in PCR, of which two were proved to be transgenic plants by southern blotting. |
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| Keywords: | Hs1 pro-1 expression vector construction particle bombardment soybean |
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