Synthesis of cDNA Pun1 gene from Capsicum frutescens L. cv. Cakra Hijau

Abstract

Capsicum frutescens L. cv. Cakra Hijau is a local cultivar that has been widely cultivated in Indonesia due to the pungency. Pungent on Capsicum is generated by capsaicin compound encoded by Pun1 gene. The sequences of Pun1 gene containing with two exons that located on the upstream and downstream, which are separated by introns in the middle. This study aimed to synthesis of cDNA of Pun1 gene from isolated total mRNA using two primers: F1/R1 (F1 5'-ATG GCT TTT GCA TTA CCA TCA-3'; and R1 5'-CTT AGC TCG AAG TGC ATC TA-3') to synthesis the exon-1 sequences and F2/R2 (F2 5'-GAA GGT GGC AGA AGA ATC AG-3'; and R2 5'-TTA GGC AAT GAA CTC AAG GA-3') to synthesis the exon-2 sequences. The cDNAs resulted from RT-PCR were visualized on 1.5% agarose gel electrophoresis. From this study we obtained a 1323 bp fragment cDNA.