Full Length Research Paper

Agrobacterium mediated transformation of annexin gene in tobacco (Nicotiana tabacum)

Thangavelu Pugalendhi¹*, Murugesan Premalatha¹ and Shreedhar g. bhat²

¹Post Graduate Department of Biotechnology, Ayya Nadar Janaki Ammal College (Autonomous), Sivakasi – 626 124, Tamil Nadu, India.

²Shreedhar Bhat’s Laboratory, 2456 Monith Complex, Bangalore-560070, India.

*Corresponding author. E-mail: marinedrug@rediffmail.com. Tel: 04562-254100.  Fax: 04562-254970.

Accepted 12 September, 2008

   Abstract

The present study involves the development of genetically engineered tobacco plants with annexin gene. The plasmid pUC 119 with the Annexin gene and pGPTV vector were isolated from the Escherichia coli. These plasmids were subjected to restriction digestion with EcoRI and Xbal where the Annexin gene is released from the pUC 119 as a linearised band was eluted from the gel. The recombinant PGPTV plasmid with the annexin gene in Agrobacterium tumefaciens MTCC 431 was mobilized and transferred to plant system through the mobilization helper plasmid pRK2013. The kanamycin resistance gene (NPT II) was used as a selective marker. The calli used for isolating the genomic DNA which was then amplified for confirmation of annexin gene. The nptII gene of 800 bp serves as a selectable marker system in plants and its amplification confirmed the presence of annexin gene in transgenic plants by PCR method.

Key words: Annexin gene, Agrobacterium tumefaciens, Nicotiana tabacum, pGPTV vector, pUC119, transgenic tobacco.