大赖草总DNA转化小麦的分子证据

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用来自大麦基因组的4个高度重复序列克隆了pHv7161、pHv7179、pHv7191和pHv7293,经地高辛和同位素2种方法标记后作为探针,对新疆大赖草(供体)、春麦761(受体)以及用大赖草总DNA通过花粉管通道转化成功的大穗转化株基因组在高度严谨条件下进行了分子杂交。结果表明,这4个探针可以探查出基因组内一种具有主串联重复单位的散在重复序列。比较受体、供体和转化株的杂交图谱,发现在转化株中出现了受体没有而供体具有的带。把上述经pHv7179探针探查出的在供体(大赖草)和转化株中共有的Hind片段进行克隆,将大赖草片段克隆命名为pLR980。经与上述4种大麦重复序列杂交证明pLR980包含了4种大麦重复序列的同源序列。用大赖草DNA克隆pLR980对供体大赖草、受体春麦761和转化株大穗小麦的基因组进行了分子杂交,结果表明pLR980在大赖草和小麦的基因组间是同源的,并且再次探查出了供体大赖草与转化株大穗小麦共有而受体没有的带。说明pLR980为大赖草总DNA通过花粉管通道法转化小麦的相关DNA片段。这为总DNA导入提供了直接的分子证据。这个转化片段已经过分离克隆,可用于进一步的研究。
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