Lee K Y, Lund P, Lowe K, Dunsmuir P
DNA Plant Technology, Oakland, California 94608.
Plant Cell. 1990 May;2(5):415-25. doi: 10.1105/tpc.2.5.415.
A single amino-acid change in the acetolactate synthase (ALS) protein of tobacco 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.
烟草乙酰乳酸合酶（ALS）蛋白中的单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中缺失的、无功能的片段，携带指定氯磺隆抗性的突变位点以及紧密连锁的新型限制性位点标记，被克隆到二元载体中。用携带该载体的根癌农杆菌转化烟草原生质体，产生了对氯磺隆有抗性的菌落。对这些菌落的DNA进行凝胶印迹分析表明，在三个转化体中，内源ALS基因与导入的T-DNA中存在的缺失ALS基因之间发生了同源重组。从这些对氯磺隆有抗性的菌落中再生出植株，在两个转化体中，对其后代的遗传分析表明，该新基因作为单个孟德尔位点进行分离。讨论了产生这些重组植物的可能模型。
