蜡梅CpPR-4基因的克隆及植物表达

Molecule Cloning and Plant Expression on CpPR-4 from Chimonanthus Praecox(L.)

在现有的蜡梅cDNA文库的基础上克隆了蜡梅病程相关蛋白4基因,并将其克隆至植物表达载体pCAM-BIA2301g上,构建了该基因的融合表达载体pCAMBIA2301g/CpPR-4,转入至大肠杆菌LBA4404后利用叶盘法转化至烟草并进行相关功能分析,通过抗病性实验及低温胁迫实验分析,转基因烟草对病毒无明显抗性,对低温不利环境有一定抗性.

Based on constructed cDNA library from Chimonanthus praecox Expressed Sequence,a gene named PR-4 has been obtained through full length sequencing of cDNA library clone,and be cloned into expression vector pCAMBIA2301g to produce a plant expression vector pCAM/CpPR-4.The recombinant plasmid is transferred into Agrobacterium tumafaciens LBA4404 by leaf disc method successfully,and to be the related function anailsis.We have conducted TMV infection test and the low temperature stress test.The test of infection of TMV virus have showed that transgenic plants are not able to resist TMV,but the transgenic tobacco has a certain resistance to the unfavorable environment of low temperature compared with wild-type tobacco.