Penyisipan Gen LEAFY Lengkap Tanaman Kakao pada Vektor Ekspresi dan Transformasi ke dalam Agrobacterium sp.

Abstract

Cacao (Theobroma cacao L.) is one of the plantations that has important role for the national economy. However, one of the problems faced in the cocoa plantations is cherelle wilt. Cherelle wilt reduces the level of productivity of cocoa. Flowering genes regulate the normal flower formation in cacao. One of the genes that play a major role in the flower formation is the LEAFY gene. Studying the flowering genes are expected to elucidate the effect of these genes to cherelle wilt. The objective of this research is to construct expression vector harbouring fulllength of LEAFY gene. The fulllength of LEAFY gene that has been inserted into the expression vector was then transformed into Agrobacterium tumefaciens AGL-0. Cloning of LEAFY gene into expression vector was carried out using the Gateway technology. The full size LEAFY gene of 1200 bp was first inserted into a donor vector and then inserted into the expression vector. Characterization of expression vector inserted with LEAFY gene was carried out using the PCR plasmid technique which resulted in a DNA band of 500 bp. Expression vector inserted with LEAFY gene was transformed into the Agrobacterium tumefaciens AGL-0. Analysis of recombinant plasmid after transformation into the Agrobacterium tumefaciens AGL-0 using PCR plasmid technique result in a DNA band of 500 bp. The conclusion of this research is that the fulllength of LEAFY gene had successfully been inserted into the expression vector with Gateway technology.