Konstruksi Gen Kitinase untuk Ketahanan terhadap Ganoderma pada Vektor Ekspresi pCambia 1303

Abstract

Efforts to increase palm oil production face problems associated with pest and diseases caused by pathogenic fungus such as Ganoderma spp. A new approach to obtain palm oil transgenic is by inserting a gene encoding antifungi compound. One of genes is chitinase gene derived from Trichoderma. The research targets are to insert chitinase gene into expression vector pCambia 1303 between the 35S Cauliflower Mosaic Virus (CaMV) promoter and the root specific promoter. The Chitinase gene was cloned into pCAMBIA 1303 expression vector followed by transformation of recombinant plasmid into Escherichia coli XL-1 Blue and transferred into Agrobacterium tumefaciens. Results showed that the chitinase gene was successfully inserted into pCambia 1303. Restriction of the recombinant DNA using restriction enzymes Nco1 and Spe1 resulted in two bands of 1200 bp which is pCambia 1303 and of 1500 bp which is the chi gene. The recombinant DNA was successfully transformed into Agrobacterium tumefaciens using competent cell of AGLO strain. The success of transformation was analyzed by restricting recombinant DNA isolated from transformant colonies using NcoI and SpeI restriction enzymes.