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油菜PEP基因的克隆及PEP反义基因的构建
引用本文:陈锦清,黄锐之.油菜PEP基因的克隆及PEP反义基因的构建[J].浙江大学学报(农业与生命科学版),1999,25(4):365-367.
作者姓名:陈锦清  黄锐之
作者单位:浙江省农业科学院原子能利用研究所!浙江杭州310021
基金项目:国家自然科学基金,浙江省重点攻关项目
摘    要:丙酮酸羧化酶( P E P)是控制油菜蛋白质/油脂含量比例的一个关键酶⒚本研究用 P C R 法扩增出了 P E P 基因片段,并将其 克隆到 p B S S K+ 的 Sm a Ⅰ位点⒚ D N A 序列分析表 明克隆片段 的长度为 530bp,其序列与报道序列相同⒚将该 P E P基因 片段反向插入 p I G121 质粒,构建了带 P E P 反义基因的超级双元载体并进行油菜的转化,目前已获得转基因植株⒚

关 键 词:丙酮酸羧化酶(PEP)  反义基因  基因克隆  双元载体  PCR

Molecular cloning and sequencing of the PEP gene from Brassia napus and the construction of the antisense PEP gene.
CHEN Jin qing,HUANG Rui zhi,Lang Chun xiu,HU Zhang hua,LIU Zhi hong.Molecular cloning and sequencing of the PEP gene from Brassia napus and the construction of the antisense PEP gene.[J].Journal of Zhejiang University(Agriculture & Life Sciences),1999,25(4):365-367.
Authors:CHEN Jin qing  HUANG Rui zhi  Lang Chun xiu  HU Zhang hua  LIU Zhi hong
Abstract:Phosphoenolpyruvate carboxylase (PEP) plays an important role in the control of the ratio of the protein/lipid content in rape ( Brassica napus ). The PEP gene fragment of B. napus was amplified with polymerase chain reaction (PCR) and then cloned into Sma I site of the vector pBS SK . The results of DNA sequence analysis indicated that the cloned fragment was 530 bp in size. In order to down regulated expression of the PEP gene, this fragment was inserted in an antisense orientation between the CaMV 35S promoter and the GUS gene of the super binary vector cassette pIG121. The antisense PEP gene was transferred into rapes ( Brassica napus ) via the Agrobacterium mediated method and the transgenic plants had been obtained.
Keywords:PEP  antisense gene  cloning  binary vector
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