KARAKTERISASI FRAGMEN 0.45 kb GEN PENGKODE STILBEN SINTASE (STS) DARI TANAMAN MELINJO (Gnetum gnemon L.); CHARACTERIZATION OF 0.45 kb GENE FRAGMENT ENCODING STILBENE SYNTHASE FROM MELINJO (Gnetum gnemon L.)

Elly Rustanti, Tri Joko Raharjo

2011 | Disertasi | PROGRAM STUDI S2 KIMIA

Characterization of 0.45 kb gene fragment encoding stilbene synthase (STS) from melinjo (Gnetum gnemon L.) has been carried out. Study was aimed to obtain nucleotide sequence of melinjo sts gene fragment. A RT-PCR (Reversed Transcriptase Polymerase Chain Reaction) amplification was performed using primers that were designed based on the amino acid conserve sequence of 10 STS protein obtained from the genebank. Two-steps RT-PCR amplification was conducted to a total RNA product isolated from the melinjo leaf. An obtained DNA fragment was purified and sequenced subsequently. The nucleotide sequence was then analyzed and compared to sts Arachis hypogaea to determine its homology to an existed sts gene. The designed primers were GGF1 (5’GCAACCGTCCTGGCAATCGC3’) and GGR1 (5’ GTTCCACCTGCGAAGCAGCC 3’) which were predicted in position 49-69 and 487-506 of sts gene respectively. The RT-PCR results in severals bands with one of them has been isolated as 0.45 kb fragment. The sequence analysis of the 0.45 kb fragment showed that the DNA sequence had 77% similarity to Arachis hypogaea sts gene. This homology was higher than the 66% similarity commonly obtained among sts genes of the various plants. Therefore the amplified fragment could be concluded as part of gene sts melinjo.

Kata Kunci : stilben synthase (sts) gene, RT-PCR, melinjo (Gnetum gnemon L.)