Isolasi dan Karakterisasi Promoter Gen AGAMOUS (TcAG) dari Kakao (Theobroma cacao L.)

Abstract

Cocoa is one of the plantation commodities that is important to the national economy. Various efforts have been made to increase cocoa production. One such efforts in the field of genetic engineering discovery for the promoter of gene that has important role in flowering and fruiting. The aim of this research to isolate the promoter of reproductive gene AGAMOUS from Theobroma cacao L. (TcAG) using Thermal Asymmetric Interlaced Polymerase Chain Reaction (TAIL PCR) method and obtain the characteristics of the promoter using a computer program online Softberry. Technique of TAIL PCR using the reverse genespecific primers TcAG position +202 (TcAG+202) (5'- GGCCACGGCTAGAGAAGACAA-3') and position +67 (TcAG+67) (5’- ATCTCGATCTTTCCCCTTCCC-3'). Forward primer AP140 provided the best results are fragment genomic size of 1200 bp. Analysis of Transcription Start Site Prediction (TSSP) of softberry program from the 5' of the fragment has proven that it has three promoter regions. The regions which are nucleotide in position 126 (TATA box at position 93), nucleotide at position 735 (TATA box at position 702) and nucleotide at position 807 (TATA box at position 774). TATA box at position 774 is predicted to be the functional binding sites of RNA polymerase because it is in the position closest to the transcription start site.