文献类型：期刊文献

中文题名：香石竹ACC氧化酶基因的克隆与植物表达载体构建

英文题名：Cloning of ACC Oxidase Gene of Carnation and Construction of Its Plant Expression Vectors

作者：余义勋[1] 张俊卫[1] 孙振元[2] 包满珠[1]

第一作者：余义勋

机构：[1]华中农业大学林学系;[2]中国林业科学研究院花卉中心

年份：2002

卷号：15

期号：3

起止页码：256-260

中文期刊名：林业科学研究

外文期刊名：Forest Research

收录：CSTPCD;;Scopus;北大核心:【北大核心2000】；CSCD:【CSCD2011_2012】；

基金：国家自然科学基金 (39970 5 32 ) ;94 8项目 (96 0 4 0 6 )资助

语种：中文

中文关键词：植物表达载体;香石竹;ACC氧化酶;基因克隆;重组载体

外文关键词：carnation ( Dianthus caryophus L.); ACC oxidase gene; cloning; sequencing; construction of plant expression vector

分类号：S681.5

摘要：根据已发表的ACO核苷酸序列 ,设计一对引物 ,以香石竹品种‘American’基因组DNA为模板 ,扩增出香石竹ACC氧化酶基因 ,将其连接到pMD18加T质粒载体上 ,通过中间载体pBluescriptSK ,构建了ACO基因正向和反向插入的植物表达载体 ,并将其导入了农杆菌。酶切检测表明该片段已插入表达载体 ,序列测定结果显示所克隆基因片段含 3个外显子 ,2个内含子 ,其中
A pair of primers were designed according to the 1 aminocyclopropone 1 carboxylic acid (ACC) oxidase gene of carnation and the primers were used to amplify the genomic DNA fragment of about 1.2 kb by polymerase chain reaction (PCR)by taking genome DNA from 'American' carnation leaves as template. The PCR product was cloned into T tailing pMD18 vector. Sequencing indicated that the ACC oxidase gene included three exons interrupted by 2 introns with identical positions as they are in tomato. ACC oxidase gene were respectively cloned into plant expression vector pMOGMON in sense and antisense orientation. Recombinant expression vectors were identified by restriction enzyme and PCR analysis. PCR indicated plant expression vectors were transferred into A. tumefaciens .