Construction of MYMIV-based gene silencing vector and its use

In this present era of ¿Genomics,¿ the genomes of many different organisms or plant species are already sequenced and some are being sequenced. As a result,a huge repository of unknown genes or genes with unknown functions has developed and ¿functional genomics¿ has to deal with the problems of such¿unknowns¿. We have constructed a novel VIGS vector,using the minimal replicon region of the DNA-A component of the geminivirus, Mung bean Yellow Mosaic India Virus (MYMIV). The vector is incorporated between the left and right border region of standard pCAMBIA vector so that the recombinant vector could be delivered into the plant following the usual agro-bacterial infection techniques. The MYMIV-based vector replicated as an episomal entity within 10-15 days past agro-infection in most of the legumes,tobacco and Arabidopsis plants. The silencing efficacy of this vector was demonstrated using GFP as a transgene and PCNA as an endogene. This vector is supposed to silence a large array of endogenes from a wide variety of plants in a fast track and cost-effective mode.