摘要
磷酸烯醇式丙酮酸羧化酶(PEPC)是控制花生蛋白质/油脂含量比例的一个关键酶。本研究利用PCR方法扩增出了PEPC基因片段,并将其克隆到pMD18-T Simple载体,序列分析表明克隆片段的长度为728bp。将该PEPC基因片段反向插入pCAMBIA1301-35S双元表达载体,构建了带PEPC反义基因的双元载体并进行花生的转化,目前已获得转基因植株,收获T1代种子。
Phosphoenolpyruvate carboxylase（PEPC）plays an important role in the control of the ratio of the protein/lipid content in peanut（Arachis hypogaea L.）.The PEPC gene fragment of peanut was amplified with polymerase chain reaction（PCR）and then cloned into the vector pMD18-T Simple.The results of DNA sequence analysis indicated that the cloned fragment was 728bp in size.In order to down regulated expression of the PEPC gene,this fragment was inserted into an antisense orientation between the CaMV 35Spromoter and the GUS gene of the binary vector pCAMBIA130135S.The antisense PEPC gene was transferred into peanut（Arachis hypogaea L.）via the Agrobacterium mediated method and the T1transgenic seeds had been obtained.
出处
《花生学报》
2013年第2期9-13,共5页
Journal of Peanut Science
基金
国家产业体系(CARS-14)
山东省自然基金(ZR2009DQ004
ZR2011CQ036
ZR2012CQ031)
国家自然基金(31000728
31100205
31200211)
山东省优秀中青年科学家科研奖励基金(BS2010NY023)
青岛市科技计划(11-2-4-9-(3)-jch
11-2-3-26-nsh
12-1-4-11-(2)-jch)
关键词
花生
PEPC基因
载体构建
遗传转化
peanut
PEPC gene
vector construction
genetic transformation
