Abstract:
This study is carried to optimize Arabidopsis transformation protocol using β-glucuronidase (GUS) gene as a reporter gene to analyse gene expression in heterologous system. The method we used to facilitate our studies is Agrobacterium mediated floral dip method with minor modifications. Floral dip method is the most popular protocol for the production of stable transformants of Arabidopsis thaliana. Agrobacterium has the ability to transfer foreign DNA into intact plant cells. In floral dip method, the developing inflorescences of Arabidopsis were dipped in the suspension of overnight grown Agrobacterium carrying the genes of interest for about 10 seconds. This dipping of inflorescences targeted the female gametes resulting in transformed seeds. The transformed seeds were then plated in vitro on a selective medium and screened for primary transformants (T0). T0 transgenic lines were then planted to get T1 followed by sowing for T2 and T3 respectively. Using this method T2*- transgenic plants, homozygous for transgene, were obtained in 10.5 months.
Page(s):
287-290
DOI:
DOI not available
Published:
Journal: Science International, Volume: 25, Issue: 2, Year: 2013