Capsicum frutescens L. is one of Chili peppers with high pungency. Capsicum frutescens has several cultivars, one of those is C. frutescens cv. Cakra Hijau. This cultivars is known resistance to pests and diseases as well. Pungency is due to the accumulation of capsaicinoids. Pun1 is an important gene responsible for pungency. The full-leght genomic sequence of Pun1 is 1897 bp, containing two exons of 738 bp and 590 bpand one intron of 348 bp in between. This study was aimed to isolate Pun1 gene that free from intron. mRNA was isolated with TriReagentâ furthermore RT-PCR method used Qiagent One-Step RT-PCR and two pairs of primer : F1/R1 (F15'-ATG-GCT-TTT-GCA-TTA-CCA-TCA-3' / R15'- CTT-AGC-TCG-AAG-TGC-ATC-TA-3') and F2/R2 (F25'-GAA-GGT-GGC-AGA-AGA-ATC-AG-3'/R25'-TTA-GGC-AAT-GAA-CTC-AAG-GA-3'). The result of this study are isolated 738 bp exon-1 and 590 bp exon 2 of Pun1 gene