Construction of Plant Expression Vector of Synthetic Bt- cry1Ac Gene for Genetic Transformation

摘要

To construct the plant expression vector containing synthetic B. thuringiensis-cry1Ac gene for crop plant transformations to resist the insect Helicoverpa armigera. A newly constructed binary vector containing the T - DNA left border, Kanamycin (kan) as marker gene, glucuronidase (uidA) reporter gene and bt-cry Ac 1 gene which transformed to A. tumefaciens by helper plasmid pRK2013, which provides tra and mob genes required to transfer the DNA. The plasmid constructed from basic vector pUC118 containing synthetic Bt-cry 1 Ac gene and pGPTV was restricted digested by EcoRI and Xbal. The digested plasmids were purified, quantified and ligated before triparental mating method of competent cell transformation. The triparental mating efficiency can be observed through back transformation of gene from A. tumefaciens to E.coli. The confirmation of Bt-cry 1 Ac gene in the construct was done by Polymerase Chain Reactions (PCR). Construction of a Bt-cry Ac1 expression vector was successful and this study will be a feasible approach for the genetic improvement of an economically important Crop plants.