Isolation 3’-end Fragment of Pun1 gene from Capsicum frutescens L. cultivar Cakra Hijau
Abstract
Pun1 gene [A1] is the one of candidate gene that responsible to determine pungency in Capsicum. In previous researches, 1 310 bp fragment of 1 671 bp Pun1 gene from Capsicum frutescens L. cv. Cakra Hijau had been isolated. The purpose of this research was to isolate of 3’-end fragment and get full length of Pun1 gene from C. frutescens L. cv. Cakra Hijau. DNA isolation was done using modified procedure. The method used to isolate the gene was PCR with a pair of primer, forward primer 5’-GAA-GGT-GGC-AGA-AGA-ATC-AG-3’ and reverse primer 5’-TTG- TTG ACC-GTA-AAC-TTC-CG- 3’. The result successfully to get 715 bp length DNA fragment. The assembly of this fragment into previous research produced a full length of 1 671 bp Pun1 gene from C. frutescens L. cv. Cakra Hijau consist of 738 bp first exon fragment, 348 bp intron fragment, and 585 bp second exon fragment.
Keywords: C. frustescens L. cv. Cakra Hijau; gene isolation; Pun1 gene.
References
[1] R. Rukmana, Usaha tani cabai rawit [Farm cayenne pepper], Kanisius, Yogyakarta,
2002.
[2] M. Materska and I. Perucka, “Antioxidant activity of the main phenolic compounds isolated from hot pepper fruit (Capsicum annuum L.),” Journal of Agricultural and Food Chemistry, vol. 53, no. 5, pp. 1750–1756, 2005.
[3] J. Lee, Y. Li, C. Li, and D. Li, “Natural products and body weight control,” North American Journal of Medical Sciences, vol. 3, no. 1, pp. 13–19, 2011.
[4] A. Mori, S. Lehmann, J. O’Kelly et al., “Capsaicin, a component of red peppers, inhibits the growth of androgen-independent, p53 mutant prostate cancer cells,” Cancer Research, vol. 66, no. 6, pp. 3222–3229, 2006.
[5] L. Koleva Gudeva, S. Mitrev, V. Maksimova, and D. Spasov, “Content of capsaicin extracted from hot pepper (Capsicum annuum ssp. microcarpum L.) and its use as an ecopesticide,” Hemijska Industrija, vol. 67, no. 4, pp. 671–675, 2013.
[6] M. R. Aluru, M. Mazourek, L. G. Landry, J. Curry, M. Jahn, and M. A. O’Connell, “Differential expression of fatty acid synthase genes, Acl, Fat and Kas, in Capsicum fruit,” Journal of Experimental Botany, vol. 54, no. 388, pp. 1655–1664, 2003.
[7] C. Stewart Jr., B.-C. Kang, K. Liu et al., “The Pun1 gene for pungency in pepper encodes a putative acyltransferase,” Plant Journal, vol. 42, no. 5, pp. 675–688, 2005.
[8] E. Blum, M. Mazourek, M. O’Connell et al., “Molecular mapping of capsaicinoid biosynthesis genes and quantitative trait loci analysis for capsaicinoid content in Capsicum,” Theoretical and Applied Genetics, vol. 108, no. 1, pp. 79–86, 2003.
[9] S. Chatterjee, M. Asakura, N. Chowdhury et al., “Capsaicin, a potential inhibitor of cholera toxin production in Vibrio cholerae,” FEMS Microbiology Letters, vol. 306, no. 1, pp. 54–60, 2010.
[10] M. Habibi, A. Madhihah Manggabarani, E. Sri Sulasmi, and D. Listyorini, “AT3 (Acyltransferase) Gene Isolated From Capsicum frutescens cv. Cakra Hijau,” Journal of Tropical Life Science, vol. 3, no. 2, pp. 83–86, 2013.
[11] A. M. Manggabarani, R. R. Juliandari, E. N. Khasna, S. Zairina, E. S. Sulasm, and D. Listyorini, “ISOLATION OF UPSTREAM AND DOWNSTREAM FRAGMENT OF ACYLTRANSFERASE (AT3) GENE FROM Capsicum frutescens L. CV. CAKRAHIJAU,” KnE Life Sciences, vol. 2, no. 1, pp. 21–, 2015.
[12] , Isolasi fargmen tengah ketiga gen pun1 dari Capsicum frutescens L. kultivar Cakra Hijau [Isolation of gene fragments middle third was one of Capsicum frutescens L. cultivar Cakra Hijau]. [Thesis]. Malang: Fakultas Matematika dan Ilmu Pengetahuan Alam (FMIPA), Universitas Negeri Malang; 2015 [in Bahasa Indonesia].