Abstract:

In order to effectively transcribe dsRNA in plants, an inducer of RNA silencing, the specific primers was designed according to the published sequence of TMV ΔMP gene and CMV ΔRep gene. MP and Rep fragment was obtained by PCR amplification respectively using pBIN-TMV ΔMP (i/r) and pBIN-CMV ΔRep(i/r) as template, then MP and Rep were concatenated and cloned into pGEM-T Easy Vector. Sense MP-Rep fragment was obtained by digesting with BamHI and Kpn I and inserted into vector pKAN-In, then called the recombinant plasmid +pKAN. Antisence MP-Rep gene cut with Xba I was linked with +pKAN to construct hairpin structure. The fragment including Intron, the sense and antisence ΔMP-Rep gene was cut by Not I, and was inserted into pART27 to construct plant transformation vector. The vector could be applied in transgenic breeding engineering for plant viral resistance.