ISOLATION OF UPSTREAM AND DOWNSTREAM FRAGMENT OF ACYLTRANSFERASE (AT3) GENE FROM <i>Capsicum frutescens</i> L. CV. CAKRAHIJAU
Capsicum frutescens cv. CakraHijau is a local cultivar that has been widely cultivated in Indonesia due to its several advantages, including its pungency. Pungent taste of Capsicum is generated by capsaicin compound encoded by AT3 gene. Recently, 404 bp fragment of AT3 gene had been isolated. This research aimed to isolate upstream and downstream fragments of AT3 gene. PCR method used two pairs of primers: F2/R2 (F2 5’-TCT CCA TGC TGA CAA CAA CA-3’, and R2 5’-CGA TGA AAG ATA GCT TGT G-3’) and F3/R3 (F3 5’-GCA TCT CTT GCA GAG AGC ATA G-3’, and R3 5’-TGT ACG CAC TCG TTG AGA CT-3’). F2/R2 primers amplified 326 bp upstream fragments, while F3/R3 primer amplified 261 bp downstream fragments. The alignment of those two fragments with one previously obtained produces a 675 bp partial sequence with 230 bp located upstream of presumed start codon. ClustalX analysis reveals that this fragment is located upper half compare to C. frutescens cv. Shuanla AT3 gene. Further primer design is necessary to obtain downstream of AT3 gene.
Keywords: Capsicum frutescens cv. CakraHijau, Capsaicin, AT3 gene
Djarwaningsih, T. 2005. Capsicum spp. (Cabai): Asal. Persebaran dan Nilai Ekonomi. Biodiversitas 6 (4): 292-296.
Rukmana, H.R. 2002. Usaha Tani Cabai Rawit. Yogyakarta: Penerbit Kanisius.
Stewart, C., M. Mazourek, M.S. Giulia, O.C. Mary, and J. Molly. 2007. Genetic Control of Pungency in C. chinensis Via the Pun1 Locus. Journal of Experimental Botany 58: 979-991.
Stewart, C., B.C. Kang, K. Liu, M. Mazourek, S.L. Moore, E.Y. Yoo, B.D. Kim, I. Paran, and M.M. Jahn. 2005. The Pun1 gene For Pungency In Pepper Encodes A Putative Acyltransferase. The Plant Journal 42: 675-688.
Stellari, G.M., M. Mazourek, and M.M. Jahn. 2010. Contrasting modes for loss of pungency between cultivated and wild species of Capsicum. Heredity 104: 460-471.
Kim, M.W., K. Shinje, K. Soohyun, and D.K. Byung. 2001. Isolation of cDNA Clones Differentially Accumulated In The Placenta of Pungent Pepper by Suppression Substractive Hybridization. Molecular Cells 11 (2): 213-219.
Habibi, M., A.M. Manggabarani, E.S. Sulasmi, and D. Listyorini. 2013. AT3 (Acyltransferase) Gene Isolation from Capsicum frutescenscv. CakraHijau. Journal of Tropical Life Science 3(2): 83 – 86.