文档介绍：中国农业科技导报 2007,9(3):66~72
Journal of Agricultural Science and Technology
水稻乙酰乳酸合成酶基因的克隆和功能分析
宋贵生,冯德江,魏晓丽,唐家斌,朱祯
(中国科学院遗传与发育生物学研究所,北京 100101)
摘要:乙酰乳酸合成酶(ALS)是植物和微生物中亮氨酸、异亮氨酸和缬氨酸合成途径的一个关键酶,该基因
在植物抗除草剂基因工程中具有重要的应用价值。利用生物信息学分析,自水稻中分离了ALS基因的cDNA和基因
组DNA序列。序列分析表明,ALS基因没有内含子,GC碱基含量高并不对称分布。软件预测ALS基因是单拷贝基
因位于第 4 号染色体上,Southern杂交验证了此结果。构建ALS基因的植物表达载体并转化烟草,转基因烟草通过
PCR-Southern验证。EXACT-RT-PCR结果表明,水稻ALS基因在转基因烟草的叶片和根部均有转录发生。利用甲嘧
磺隆进行除草剂抗性测试,和对照烟草相比转基因烟草除草剂抗性高达 50 mg·L-1。
关键词:乙酰乳酸合成酶(ALS);EXACT-RT-PCR;抗除草剂;SAGE(基因表达系列分析)

Isolation and Functional Analysis of Rice Acetolactate-synthase (ALS)
SONG Gui-sheng, FENG De-jiang, WEI Xiao-li, TANG Jia-bin, ZHU Zhen
(Institute of ics and Developmental Biology, Chinese Academy of Science, Beijing 100101, China)
Abstract: Acetolactate-synthase (ALS) is a key enzyme in biosynthetic pathway for valine, leucine, and isoleucine in
plant and anism. It is important for plant insect-resistant ic engineering. The cDNA sequence and genome
DNA sequence of ALS gene were isolated. Sequence analysis indicated that ALS gene has no introns and its GC base
content is asymmetrical. Software analysis result speculated that ALS has one copy locating on chromosome 4, which was
confirmed by southern blotting. ALS plant expression vectors have been constructed in o and ic o
was confirmed by PCR southern blotting. EXACT-RT-PC