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水稻T-DNA插入群体的建立及侧临序列的获得与分析

DOI: 10.7668/hbnxb.2011.01.007, PP. 31-36

Keywords: T-DNA,插入突变,转基因水稻,TAIL-PCR

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Abstract:

采用农杆菌介导转化方法,将携带有GUS基因1301质粒转入日本晴水稻品种中,建立水稻T-DNA插入群体5200个。研究表明:通过PCR和SouthernBlot分析证明了再生植株为转基因植株;通过改进热不对称交错PCR(TAIL-PCR)方法,已成功地分离并测定了437个株系的侧翼序列;通过序列同源性分析表明,有147个序列只包含有载体序列,有214个序列中包含有与T-DNA右边界相邻的水稻基因组序列;通过水稻边界序列与网上水稻数据库中基因组序列比对,将3个突变体的T-DNA插入位点定位于水稻特定染色体上。

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