中国生物工程杂志

Tobacco mosaic virus (TMV) RNA and Cucumber mosaic virus (CMV) were extracted and sequenced, respectively. The full-length sequence clones of their coat proteins were therefore obtained using reverse transcription method; the target coding region sequence obtained using forward and reverse primers, was later subcloned into pUCCRNAi, then transferred into pCAMBIA2300-35S-OCS expression vector. Utilizing Agrobacterium tumefaciens LBA4404 infected Nicotiana tabacum K326, Then got three transform materials which containing interference sequence of coat protein gene of TMV or CMV, it has been proofed using molecular method that they have been introduced into Nicotiana tabacum successfully and we also analyzed the mRNA expression differences using real-time quantitative PCR. These data established that the transformed tobacco strains showed resistance to TMV and CMV improved remarkably.