Reza MOHAMMADHASSAN1, Kasra ESFAHANI2*, Bahareh KASHEFI3
1College of Agriculture, Damghan Branch, Islamic Azad University, Damghan, IRAN
2National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, IRAN
3College of Agriculture, Damghan Branch, Islamic Azad University, Damghan, IRAN
Corresponding Author: firstname.lastname@example.org, Phone: +982144787451
Abstract. Binary vectors are widely used in Agrobacterium gene transfer in plants. Although new plant expression vectors have been designed, pBI121–based vectors are still more common. The availability of fewer numbers of restriction enzyme sites is the most important drawback in pBI121. In this article, two new vectors–based on pB121, pBI1215+1, and pBI121GUS–6–were introduced. The construction of new vectors was confirmed by PCR and digestion pattern analysis. Furthermore, the reporter gene (gus) was cloned in these vectors. The T–DNA transformation ability of the new vectors and pBI121 (as the control sample) to tobacco via Agrobacterium tumefaciens strain LBA4404 was evaluated. Transgenic plants were regenerated with BAP and NAA in a selective medium. Thereafter, plant DNA was extracted and successful gene transformation was confirmed by PCR analysis. The GUS assay also confirmed gene expression in transgenic leaves. The results indicate that the vectors are totally efficient in plant transformation.
Keyword: Agrobacterium–mediated transformation, binary vector, Plant expression vector, pBI121, pBI1215+1, pBI121GUS–6