Isolasi Promotor Gen LFY pada Tanaman Theobroma cacao L. dengan Teknik Genome WalkingTM

Abstract

Cacao is one of the main agricultural commodities in Indonesia. Unfortunately, there are many obstacles faced by Indonesian cacao farmers, which results in the low cacao productivity. A major obstacle is the cherelle wilt disease. This disease is affected by the central flowering gene (gene LFY), whose expression level is regulated by its promoter. The objective of this research is to isolate the promoter of gene LFY, to then be cloned into its cloning and expression vectors. The promoter isolation was done from a young cacao leaf. The DNA in the leaf was extracted first, and then the promoter was isolated using Genome WalkingTM technique. Subsequently, the sequencing method and Softberry software were utilized to determine the promoter’s nucleotides. The result, which was then confirmed by TATA box, proved the existence of gene LFY promoter fragment. A fragment of 1650 bp size was then cloned into cloning and expression vectors: pGEM-T Easy and pCambia 1303. Next, the plasmid of the recombinant DNA was transformed into Agrobacterium tumefaciens using the AGLO strain. The conclusion of this research is that the gene LFY promoter of Theobroma cacao L. can be isolated using the Genome WalkingTM method, enabling more information on the occurence of cherelle wilt on cacao plants.