利用Cre/loxP重组系统构建甜菜碱合成酶多基因表达载体  

Construction of multi-gene expression vector encoding enzymes for glybet synthesis by using Cre/LoxP recombination system

作　　者：王亮[1] 苏乔[1] 安利佳[1] 

机构地区：[1]大连理工大学环境与生命学院生物科学与工程系,大连116023

出　　处：《高技术通讯》2007年第7期749-754,共6页Chinese High Technology Letters

基　　金：863计划（JY03-B-18）资助项目.

摘　　要：通过使用一套基于Cre/loxP重组系统的植物多基因表达载体构建系统,将辽宁碱蓬（Suaeda liaotungesis kitag）的胆碱单加氧酶（CMO）基因、甜菜碱醛脱氢酶（BADH）基因以及烟草的核基质结合区（MAR）序列构建到同一表达载体上,得到可直接用于农杆菌转化的植物表达载体pYLTAC747H-MAR-BADH-CMO-MAR.该甜菜碱合成酶多基因表达载体的成功构建为进一步进行植物的遗传转化,以有效提高转基因植株的耐盐性提供了实验基础.实验中,用热激法替代了电击法进行质粒的大肠杆菌转化,并去掉了透析等步骤,简化了构建过程.By using Cre/LoxP recombination system, the choline monooxygenase （CMO） gene and betaine aldehyde dehydrogenase （BADH） gene derived from Suaeda liaotungensis, and the matrix attachment region （MAR） sequences isolated from tobacco were inserted into a multi - gene plant expression vector to obtain pYLTAC747H- MAR - BADH- CMO - MAR. The construction of multi - gene expression vector encoding enzymes for glybet synthesis will contribute to further genetic manipulation aimed at improving the salt tolerance of engineered plants. To simplify the approach, the electroporation method was replaced by the heat - shock method for plasmids E. coli transformation and the procedures such as dialysis etc. were eliminated.

关 键 词：CRE/LOXP重组系统 甘氨酸甜菜碱 CMO BADH MAR 多基因表达载体 

分 类 号：Q782[生物学—分子生物学]