Konstruksi promotor gen leafy kakao pada vektor ekspresi dengan sistem gateway dan transformasi ke Agrobacterium sp.

Abstract

Cacao (Theobroma cacao L.) is one of the plantations with a good prospect because the world demand of cocoa is expected to be growing. The increased demand of cacao is contributed with a major problems related to the cultivation is cherelle wilt. Cherelle wilt in cocoa is a physiological disease that cause by the abnormalities of the genes who is responsible to the flowering process. One of the genes that play a major role in the flower formation is the LEAFY gene and LEAFY gene promoter is expected to activate LEAFY genes. The objective of this research is to construct three size of LEAFY gene promoter (1041 bp, 702 bp, and 349 bp) into the expression vector with gateway system by knowing the size of an effective related to the gene expressed. LEAFY gene promoter in size 1041 bp, 702 bp, and 349 bp to be inserted into a donor vector (pDONRTM221) and then into the specific destination vector for promoter. The recombinant plasmid has been analyzed after transformation into the Agrobacterium tumefaciens strain AGL-O give a result in a DNA band about 8002 bp, 7663 bp, and 7310 bp. The conclusion of this research by the three measures of LEAFY gene promoter, which is 1041 bp, 702 bp, and 349 bp have been successfully inserted into the expression vector with the gateway system and transformed into Agrobacterium tumefaciens strain AGL-O then ready to be transferred into the plant.