甘蓝型油菜fad2基因片段的克隆和反义表达载体的构建  

Cloning of a sequence encoding fad2 from Bassica napus and constructing of antisence fad2 expression vector

作　　者：熊兴华[1] 官春云[1] 李栒[1] 王学军[1] 周小云[1] 李家洋[2] 

机构地区：[1]湖南农业大学作物基因工程湖南省重点实验室,湖南长沙410128 [2]中国科学院遗传研究所,北京100101

出　　处：《中国油料作物学报》2002年第2期1-4,共4页Chinese Journal of Oil Crop Sciences

基　　金：湖南省科技厅资助项目 (97JKY10 0 5 )

摘　　要：从甘蓝型油菜湘油 15号基因组中分离总DNA ,以此为模板进行多聚酶链式反应 (PCR)。PCR产物与克隆载体 pGEM -TEasy连接 ,经NotI酶切后插入 pBluescriptIISK +。序列测定表明 ,PCR产物为 6 5 4bp的DNA片段 ,fad2基因片段只朝 pBluescriptIISK +的T3-T7方向插入 ,经BamHI和SacI酶切后插入表达载体质粒pBI12 1相同的酶切位点 ,构建反义 fad2表达载体。The DNA fragment of fad2 gene was amplified from Xiangyou No.15 genomic DNA by polymerase chain reaction. The PCR products were cloned into pGEM-T Easy, the 650bp fragment of recombinant pGEM-T Easy digested by NotI were cloned into pSK+and then sequenced. The results indicated that the cloned fragment contained 654 nucleotides, and share 100% of homology in comparison part with the reported data. The 654bp fragment of recombinant pBluescriptIISK+digested by BamHI and SacI were cloned into plant expression vector pBI121.

关 键 词：FAD2基因 反义表达载体 甘蓝型油菜 PCR 基因克隆 

分 类 号：S565.4[农业科学—作物学]