طراحي و ساخت چهار حامل پلاسميدي نوتركيب مناسب براي انتقال ژن هاي كيتيناز، گلوكاناز و Bt به گياهان

Design and construction of four recombinant plasmid vectors containing chitinase, glucanase and BT genes, suitable for plant transformation

Four plasmid vectors that are suitable for plant transformation were constructed by preparation of gene cassettes for p-1, 3-glucanase from barley, chitinase from bean and crylA (b) from Bacillus thuringiensis (BT). Each of these genes was cloned under the control of the CaMV35S promoter and the Nos terminator in pBTl21 binary vector. Chitinase and [3-1,3- glucanase genes that encode antifungal proteins, can hydrolyzes p-1,3- glucans and chitin, respectively, which are major components of the cell walls of many pathogenic fungi and increase plants resistance against fungal infection. CrylA (b) gene also encodes an insecticide protein that is generally active against lepidopteran insects. pBI-G/i< and pBI-C/n recombinant plasmid vectors were constructed via cloning of p-1,3- glucanase and chitinase genes, respectively, instead of the gus gene in T-DNA region of pBI 121 vector. Construction of pBl-C/i/G/w recombinant plasmid vector was performed by means of cloning both of the complete chitinase and glucanase gene cassettes in pBI 121 vector, with the intention of production synergistic effects against fungal infection, pB\-ChiBt recombinant plasmid vector containing both of the complete chitinase and Bt gene cassettes was also constructed in order to simultaneous plants resistance to fungal and pest pathogens in a single transformation event. These four recombinant plasmid vectors can be useful for plant transformation via Agrobacterium and other transformation approaches such as biolistic.