Construction of Binary Vector of Cucumber Mosaic Virus Coat Proteingene

The coat protein gene of cucumber mosaic virus was as target gene;and the E.coli DH5α was as the host bacteria of the intermediate vector for vitro operation;and the Agrobacterium tumefaciens LBA4404 was as the host bacteria of the final binary vector in this paper.The target gene was connected to the pMD18-T simple Vectoer after RT-PCR,than the recombinant plasmid was transformed into E.coli by freeze-thaw method,which was insert into expression vector restriction sites after digested with restriction endonuclease.The new recombinant plasmid DNA was transformed into the Agrobacterium bacteria which contain only helper plasmid by the freeze-thaw method.Than the construction of binary vector of target gene was completed.