Journal of Environmental Biology

Detection and quantitation of GNA -transgene in

GM rice using real time PCR

Vibha Pandey¹, K. V. Rao², P.K. Shukla³, U.N. Dwivedi⁴ and Sudhir K. Goel^1*

¹CSIR, Indian Institute of Toxicology Research Lucknow-226 001, India

²Centre for Plant Molecular Biology, Osmania University, Hyderabad-500 007, India

³Department of Botany, Brahmanand P.G. College, Kanpur-208 001, India

⁴Department of Biochemistry, Lucknow University, Lucknow- 226 007, India

*Corresponding Author E-mail: sudhir.ji@gmail.com

 Publication Data

Paper received:

12 April 2012

Revised received:

08 May 2013

Accepted:

03 June 2013

Abstract

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 R² value of 0.972. As low as one copy number could be detected using the developed protocol.

 Key words

GMO, GNA, Real time PCR, SPS, SYBR green

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