Isolation of PRI gene from Tobacco genome and its cloning with PRII & PRIII genes

Raoufi, A. (2010) Isolation of PRI gene from Tobacco genome and its cloning with PRII & PRIII genes. Masters thesis, University of Zabol.

Isolation of PRI gene from Tobacco.pdf

Download (61kB) | Preview


With respect to the amount of damages resulting from fungal diseases, applying biotechnological technics for producing resistant plants is of a high significance. Complex expression of pathogenesis-related proteins induces multiple gene resistant followed by stability of resistance and also developing resistance toward a wide range of fungal diseases. In the present study, with the purpose of isolating the gene, constructing multiple vectors including 3 important group of pathogenesis-related proteins PR1, PR2 and PR3 were carried out. In this study, after designing specific primers, 2 genes with different characteristics from PR1 family were isolated from tobacco genome. After the validation test, we cloned the genes under expression of 35S and Nos terminator in the pB121 binary vector. In the next stage, cloning of two other important anti-fungal genes under control of dependent regulatory regions of T-DNA in the form of symmetric an asymmetric with PR1 in several steps were3 carried out. In order to transform the mentioned genes to monocotyledons, pIPKb010 vector were used. After adding Kozak sequence to the beginning of the gene, the whole gene cassette was constructed. Chitinase and Glucunase genes were placed in pIPUbi and after that the kPRP-1 gene were put in the middle of the mentioned gene cassette. It is expected that the three constructed vectors in this study will put up the resistance against a wide range of fungal diseases in monocot and dicot plants as well as it will show potential resistance towards the bacteria and also abiotic and biotic stresses. Eventually the constructs can be used for gene transformation by agrobacterium and biolistic gun. One of the constructs was used to transform tomato plants. Molecular analysis showed the presence of three Chitinase, gluconase and PR1 transgenes.

Item Type: Thesis (Masters)
Uncontrolled Keywords: Pathogensis related protein, gene cloning, Chitinase, Glucanase, PR1, kozak segment
Subjects: S Agriculture > SB Plant culture
Depositing User: admin admin1 admin2
Date Deposited: 25 Jun 2016 05:27
Last Modified: 25 Jun 2016 05:27

Actions (login required)

View Item View Item