HOMOLOGOUS RECOMBINATION IN PLANT-CELLS AFTER AGROBACTERIUM-MEDIATED TRANSFORMATION
HOMOLOGOUS RECOMBINATION IN PLANT-CELLS AFTER AGROBACTERIUM-MEDIATED TRANSFORMATION
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DOI：
10.1105/tpc.2.5.415
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发表时间：
1990-05-01
期刊：
影响因子：
11.6
通讯作者：
DUNSMUIR, P
中科院分区：
文献类型：
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作者：
LEE, KY;LUND, P;DUNSMUIR, P
烟草乙酰乳酸合成酶（ALS）蛋白中的一个氨基酸变化赋予了对除草剂氯磺隆的抗性。从乙酰乳酸合酶基因，携带突变位点指定绿磺隆抗性加上紧密相连的新的限制性位点标记，删除，非功能性片段克隆到一个二元载体。用携带该载体的根癌农杆菌转化烟草原生质体，获得了抗绿磺隆的菌落。来自这些菌落的DNA的DNA凝胶印迹分析表明，在三个转化体中，内源性ALS基因和引入的T-DNA中存在的缺失的ALS基因之间发生了同源重组。从这些抗绿磺隆的菌落中再生植株，并且在其中两个转化体中，对它们后代的遗传分析表明，新基因分离为单个孟德尔基因座。这些重组植物的产生可能的模型进行了讨论。
A single amino-acid change in the acetolactate synthase (ALS) protein of tobacco [Nicotiana tabacum] confers resistance to the herbicide chlorsulfuron. A deleted, nonfunctional fragment from the acetolactate synthase gene, carrying the mutant site specifying chlorsulfuron resistance plus a closely linked novel restriction site marker, was cloned into a binary vector. Tobacco protoplasts transformed with Agrobacterium tumefaciens carrying this vector yielded chlorsulfuron-resistant colonies. DNA gel blot analysis of DNA from these colonies suggested that in three transformants homologous recombination had occurred between the endogenous ALS gene and the deleted ALS gene present in the incoming T-DNA. Plants were regenerated from these chlorsulfuron-resistant colonies, and in two of the transformants, genetic analysis of their progeny showed that the novel gene segregated as a single Mendelian locus. Possible models for the generation of these recombinant plants are discussed.