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IDENTIFIKASI GEN HOUSEKEEPINGPADA DNA GENOM IKAN GURAME (Osphronemusgouramy)
Gen housekeeping merupakan gen pengatur fungsi selular basal pada organisme hidup yang secara konstitutif diekspresikan terus menerus dalam kondisi apapun. Oleh karena sifat tersebut, gen housekeepingdigunakansebagaikontrol internalpadaanalisisekspresi gen lain,dan merupakan instrumental untukkalibrasi di banyakaplikasibioteknologidanpenelitiangenom. Namun informasi genetik mengenai gen housekeeping pada ikan gurame (Osphronemus gouramy) sangat minim, sehingga penelitian ini bermaksud untuk mengisolasi dan mengidentifikasi gen-gen housekeeping pada ikan gurame dengan merancang primer degenerate yang dapat mengamplifikasi gen housekeeping pada DNA genom ikan gurame. Primer degenerate dirancang berdasarkan konsensus sikuen gen dari beberapa ikan Infraclassis Teleostei. Terlebih dahulu kandidat gen housekeeping dipilih berdasarkan studi literatur, dan diperoleh tujuh kandidat gen 18S rRNA, ODC1, HPRT1, TBP, UBC, G6PD, dan TAF2. Perancangan primer dilakukan secara online pada laman Primaclade. Amplifikasi gen housekeeping pada ikan gurame dilakukan menggunakan metode nested PCR yang mana terdapat dua set pasangan primer terdiri dari outer-primer dan primer nested. Dua primer yang mampu mengamplifikasi gen housekeeping pada DNA genom ikan gurame dilakukan sequencing untuk mengetahui urutan sikuen gen. Hasil identifikasi gen 18S rRNA dan ODC1 dari ikan gurame (Osphronemus gouramy) berdasarkan analisis BLAST dan pohon filogenetik menunjukkan semua gen yang diperoleh sesuai dengan gen target yang diharapkan dan mampu diekspresikan pada ikan gurame. Analisis BLAST pada pasangan primer spesifik yang dirancang menunjukkan gen 18S rRNA dan ODC1 spesifik pada ikan gurame;---Housekeeping genes are regulating basal cellular function genes in living organisms that are assumed constitutively to be expressed continuously under any conditions. Because of these properties, housekeeping genes are used as internal controls in the analysis of other gene expressions, and are instrumental for calibration in many applications of biotechnology and genome research. Inadequacy of genetic information about housekeeping genes in goramy (Osphronemus gouramy), a study was conducted to isolate and identification housekeeping genes in goramy by designing degenerates primer pair that can amplify housekeeping genes in the goramy genome. The degenerate primer pair was designed based on the consensus of the gene sequences of several Teleostei fish. Firstly, housekeeping gene candidates were selected based on literature studies, and seven candidates of genes 18S rRNA, ODC1, HPRT1, TBP, UBC, G6PD, and TAF2 were obtained. The primer design has been done online on the Primaclade page. Amplification of housekeeping genes in goramywere using the nested PCR method which has two sets of primer pairs consisting of outer-primers and nested primers. Only two primers capable of amplifying housekeeping genes in goramy genom. The results of 18S rRNA and ODC1 genes identification from goramy (Osphronemus gouramy) based on BLAST analysis and phylogenetic trees showed that all genes obtained were in accordance with the expected target genes and were able to be expressed in goramy. BLAST analysis on the specific primer pairs shows that the 18S rRNA and ODC1 genes specific to goramy.
Perpustakaan UPI, Universitas Pendidikan Indonesia
IDENTIFIKASI GEN HOUSEKEEPINGPADA DNA GENOM IKAN GURAME (Osphronemusgouramy) (Text)
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