Transformation of the Rapeseed (Brassica napus L.) with Firefly Luciferase Gene

Document Type : Research Paper

Authors

1 M.Sc graduate student Plant Breeding, Department of Plant Breeding, University of Tarbiat Modares

2 Associate Professor in Department of Plant Breeding and Biotechnology University of Tarbiat Modares

3 Associate Professor in Department of Biochemistry University of Tarbiat Modares

4 M.Sc graduate student Biotechnology, Department of Biotechnology, University of Tarbiat Modares

Abstract

Recent advances in molecular biology and plant biotechnology have shifted the concept of using crops as a food source to recruiting them as a bioreactor for the production of therapeutic recombinant proteins. "Molecular Farming" refers to producing valuable industrial enzymes and pharmaceutical proteins in plants through genetic engineering. Firefly Luciferase enzyme is one of the most important industrial enzymes being widely used in the various fields of Biotechnology and Cell and Molecular Biology, particularly in the detection rate of ATP to determine microbial pollution, the cancer identification kits, screening drugs, bioassay, sequencing of DNA (Pyrosequencing) and enzyme measuring. Luciferase gene has many applications, it is also ideal as a reporter gene in genetic engineering in order to optimize gene transfer systems. This study was done to transfer Iranian Firefly Luciferase gene (Lampyris turkestanicus) to Canola. In this study, Firefly Luciferase gene (luc) under the S35 promoter cauliflower virus (CaMV 35S) transferred to Agrobacterium strain LBA4404. Transformation was confirmed by Colony PCR with specific primers. recombinant plasmid bearing Firefly Luciferase genes (luc), was transferred to Canola by Agrobacterium-mediated transformation method. Cotyledon explants of Canola (cultivar PF4570/91) were used for transformation. For determining canola tolerance threshold, different concentrations of Hygromycin were analyzed. The transformed plants were screened on MS medium containing 7.5 mg.L-1 Hygromycin and 200 mg.L-1then transferred to regeneration and rooting medium. PCR analysis of transgenic palnts verified the presence of Firefly Luciferase (Lampyris turkestanicus(gene.

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Volume 3, Issue 1 - Serial Number 1
November 2012
Pages 9-16

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