Construction and Transformation of the Plant Expression Vector Carrying MsChiIGene of Tobacco

In order to improve the performance of resistance to root rot of alfalfa,by the method of the restriction endonuclease(NcoⅠ and SpeⅠ)double-enzyme digestion,the recombinant plasmid MsChipMD18-T and expression vector PCAMBIA1302from double-enzyme digestion were linked directionally with the segments of MsChiⅠgene to construct the plant expression vector MsChi-pCAMBIA1302,which was introduced into A.grobacteriumstrain LBA4404by means of rapid frozen thaw method.The results of PCR amplification and restriction digestion showed that the fragment of MsChiⅠ gene was introduced into pCAMBIA1302 plasmid and the expression vector MsChi-pCAMBIA1302 was transferred into A.grobacterium successfully.By PCR and RT-PCR preliminary testing,six transgenic lines were obtained,this shows that the objective gene had been successfully integrated into the tobacco genome.