LeCOP1LIKE基因的克隆、反义构建及微型番茄的转化  

CLONING,ANTISENSE CONSTRUCTION AND TOMATO TRANSFORMATION OF LECOP1LIKE GENE

作　　者：崔孟祥[1] 李鹏丽[1] 刘仲齐 金凤[1] 常立[1] 冯晓星[1] 王宁宁[1] 

机构地区：[1]南开大学生命科学学院植物生物学与生态学系,天津300071 [2]天津农业生物技术研究中心,天津300192

出　　处：《分子细胞生物学报》2007年第5期329-338,共10页Journal of Molecular Cell Biology

基　　金：天津市自然科学基金项目(5YFJZJC00800和05YFGHHZ00600)资助.~~

摘　　要：本文报告利用EST筛选结合RT-PCR的方法从番茄中克隆了LeCOP1LIKE基因的1 060 bp cDNA片段,并利用其非保守域构建反义RNA表达载体。利用农杆菌介导法,将LeCOP1LIKE基因的反义RNA表达载体转入微型番茄Micro-Tom,获得了10株反义LeCOP1LIKE转基因微型番茄。RT-PCR分析表明其中4个转基因株系中的LeCOP1LIKE表达被显著抑制,并发现抑制LeCOP1LIKE基因的表达导致转基因番茄的株高下降、叶片的叶绿素含量提高、果实中番茄红素含量增加。并且明显抑制转基因种子的发育。这些实验结果证明了LeCOP1LIKE基因为番茄发育过程中光形态建成的抑制因子。Here reported a 1060 bp cDNA cloning of LeCOP1LIKE gene by EST probing and RT-PCR method. In order vector was transformed into to characterize Micro-Tom via function of this gene, a LeCOP1LIKE antisense expression Agrobacterium-mediated transformation method and 10 independent transgenic lines were obtained. RT-PCR analysis showed that the expression of LeCOP1LIKE gene was evidently repressed in 4 lines of them. The transgenic plants were much shorter than their wild type control, their chlorophyll content was increased but the seed development was obviously suppressed. All these results suggested that the cloned LeCOP1LIKE gene was a negative regulator of photomorphogenesis in tomato.

关 键 词：反义RNA表达载体 LeCOP1LIKE基因 微型番茄Micro-Tom 农杆菌介导的遗传转化 

分 类 号：S641.2[农业科学—蔬菜学] Q943.2[农业科学—园艺学]