Detection and quantitation of GNA -transgene in GM rice using real time PCR

摘要

An efficient system for detection and quantitation of the Galanthus nivalis agglutinin "GNA" transgene, with reference gene Sucrose Phosphate Synthase (SPS), in GM rice, a SYBR green based real time PCR assay was optimized. Snowdrop lectin gene product(GNA) proved to be toxic towards the pest "brown plant hopper" (BPH) and green leaf hopper (GLH) feeding on rice. The transgenic rice line developed employing GNA and non transgenic control variety (Chaitanya) was used in this study. The DNA isolated from transgenic rice was serially diluted to create a standard curve of "GNA" gene and regression analysis yielded R~2 value of 0.972. As low as one copy number could be detected using the developed protocol.