CMO与BADH双基因表达载体构建及在烟草中的表达  

Construction of CMO-BADH Doublegene Expression Vectorand Its Expression in Tobacco

作　　者：刘君[1] 韩烈保[1] 陈其军[2] 信金娜[1] 何宇锋[1] 李雪[1] 

机构地区：[1]北京林业大学草坪研究所,北京100083 [2]中国农业大学,北京100094

出　　处：《中国生物工程杂志》2006年第8期5-9,共5页China Biotechnology

基　　金：国家转基因植物研究与产业化专项资助项目(J2002B006);国家"863"计划资助项目(2001AA244082);"教育部跨世纪优秀人才支持计划"资助项目

摘　　要：目的将甜菜碱合成关键酶CMO与BADH基因构建到同一表达载体中。方法用转基因方法将该表达载体导入植物体内,完善植物体内的甜菜碱合成途径,提高植物的抗旱性和耐盐性。以pC1303质粒为基础,构建了均由35S启动子驱动的CMO基因和BADH基因的植物双基因表达载体pC35SC35SB1303。利用冻融法将其导入农杆菌LBA4404中,通过农杆菌介导法分别将CMO基因、BADH基因以及该双基因表达载体导入烟草中。结果PCR检测和Northern杂交分析表明,外源基因已整合到受体植物基因组中并正常表达。结论对转基因植株及对照植株甜菜碱含量的检测结果表明,转双基因植株的甜菜碱含量明显高于转BADH基因植株、转CMO基因植株及对照植株。The objective is to perfect the approach of synthesizing lycine in plants in order to advance the resistance of drought and salt by introducing a vector which including the key enzyme CMO gene and BADH gene to the plants by transgenic methods. A doublegene plant expression vector, pC35SC35SB1303 were reconstructed based on PC1303 plasmid by CMO and BADH which were both driven by 35S promoter. It was introduced into Agrobacterium tumefaciens strain LBA4404 by freezethaw method. Transgenic tobacco plants recovered from leaf discs applying Agrobacteriummediated gene transfer carrying BADH, CMO and the doublegene vector were detected by PCR analysis and Northern blotting respectively which conclude that the foreign gene had consistented with the recipient plants＇ genome and expresses normally. The analysis of the content of lycine in transgenic plants and blank plants showed that it was higher in the doublegene vector transgenic plants than those transferred the single gene BADH or CMO and the blank plants either.

关 键 词：双基因表达载体 CMO BADH 甜菜碱含量 

分 类 号：Q789[生物学—分子生物学]