Karakterisasi Gen Major Royal Jelly Protein 2 (mrjp2) pada Lebah Apis cerana dan Apis mellifera Asal Indonesia

Abstract

Gen Major Royal Jelly Protein 2 (mrjp2) dapat dijadikan pembeda molekular lebah A. cerana dan A. mellifera. Sekuen DNA gen mrjp2 A. cerana dan A. mellifera di dalam GenBank terbatas dan belum pernah dilaporkan berasal dari Indonesia, sehingga diperlukan karakterisasi gen mrjp2 untuk sekuen DNA tersebut. Penelitian ini bertujuan mengarakterisasi dan menganalisis bioinformatika ekson-intron gen mrjp2 lebah Apis cerana dan Apis mellifera asal Indonesia. Metode yang digunakan meliputi ekstraksi, amplifikasi, sequencing DNA dan analisis bioinformatika. Sampel A. cerana asal Bogor dan A. mellifera asal Boyolali diamplifikasi secara berturut-turut pada Temperatur annealing (Ta) 47 oC dan 50 oC. Panjang sekuen DNA A. cerana dan A. mellifera yang didapatkan secara berturut-turut 579 pb (ekson 1-3) dan 76 pb (intron 2). Kromatogram sampel A. mellifera menunjukkan puncak ganda sehingga analisis selanjutnya dilakukan hanya pada sampel A. cerana. Panjang asam amino putative sampel A. cerana adalah 112 aa. Variasi nukleotida sampel A. cerana lebih banyak pada ekson 2 dibandingkan ekson 3. Topologi pohon filogenetik sampel A. cerana membentuk satu kluster besar dengan mrjp famili, hasil BLAST-X dan BLAST-P asam amino A. cerana 99.1% homolog dengan A.cerana asal Korea sehingga dapat digolongkan sebagai sekuen A. cerana gen mrjp2.

Major Royal Jelly Protein 2 (mrjp2) gene can be used as a molecular marker to discriminate Apis cerana and A. mellifera. DNA sequences of the mrjp2 gene A. cerana and A. mellifera were limited in GenBank and samples from Indonesia have never been reported; therefore DNA characterization and bioinformatics analysis were needed. This research was aimed to characterized and bioinformatics analyzed exon-intron mrjp2 gene of Indonesian A. cerana and A. mellifera. Methods used were DNA extraction, amplification, sequencing, and bioinformatics analysis. Apis cerana from Bogor and A. mellifera from Boyolali were amplified at Temperature annealing 47 oC and 50 oC, respectively. The length of A. cerana and A. mellifera DNA sequences were 579 bp (exon1-3) and 76 bp(intron 2), respectively. Chromatogram result showed A. mellifera formed many double-peak; thus, it could not be included in the further analysis. The putative amino acid of A. cerana the mrjp2 gene was 112 amino acids. Nucleotide variation in A. cerana greater in exon 2 than exon 3. Phylogenetic tree construction showed that A. cerana formed a big cluster with the mrjp family, the BLAST-X and BLAST-P showed 99.1% homolog with Korean A. cerana. Thus, the sequences were concluded as A. cerana mrjp2 gene.