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黄瓜磷脂酶D基因片段克隆及其反义表达载体的构建
引用本文:杜栋良,王秀峰,史庆华,杨凤娟,孙晓琦,徐慧妮.黄瓜磷脂酶D基因片段克隆及其反义表达载体的构建[J].山东农业大学学报(自然科学版),2009,40(1).
作者姓名:杜栋良  王秀峰  史庆华  杨凤娟  孙晓琦  徐慧妮
作者单位:山东农业大学园艺科学与工程学院,山东,泰安,271018
摘    要:采用RT-PCR方法,从新泰密刺黄瓜的幼叶中得到长为1018 bp的cDNA片段,编码339个氨基酸,与GenBank中甜瓜磷脂酶D基因相比核苷酸同源性为96%,氨基酸同源性为97%.克隆片段经双酶切消化,反向插入到植物表达载体pBI121的CaMV 35S启动子和NOS终止子之间,构成磷脂酶D基因的反义表达载体.

关 键 词:黄瓜  磷脂酶D  反义表达载体

MOLECULAR CLONING OF A CUCUMBER CDNA FRAGMENT ENCODING A PHOSPHOLIPASE D(PLD)AND CONSTRUCTION OF ANTISENSE EXPRESSION VECTOR
DU Dong-liang,WANG Xiu-feng,SHI Qing-hua,YANG Feng-juan,SUN Xiao-qi,XU Hui-ni.MOLECULAR CLONING OF A CUCUMBER CDNA FRAGMENT ENCODING A PHOSPHOLIPASE D(PLD)AND CONSTRUCTION OF ANTISENSE EXPRESSION VECTOR[J].Journal of Shandong Agricultural University,2009,40(1).
Authors:DU Dong-liang  WANG Xiu-feng  SHI Qing-hua  YANG Feng-juan  SUN Xiao-qi  XU Hui-ni
Institution:College of Horticulture Science and Engineering;Shandong Agricultural University;Tai'an 271018;China
Abstract:A cDNA fragment of a phospholipase D(PLD)was amplified by RT-PCR from young leaves of cucumber (Cucumis sativus L.).DNA sequencing showed that the cloned fragment shared highly homogeneity to the cDNAs of phospholipase D from muskmelon.The cloned cDNA of phospholipase D was further introduced into binary vector pBI121 in a reverse orientation with its upstream promoter(CaMV 35S),giving an expressing plasmid containing the antisense phospholipase D gene.
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