KONSTRUKSI VEKTOR EKSPRESI GEN HORMON PERTUMBUHAN LELE DUMBO (Clarias sp.) UNTUK PRODUKSI IKAN LELE LOKAL (Clarias batrachus) TRANSGENIK

Ibnu Dwi Buwono, Nono Carsono, M. Untung Kurnia

Abstract


Penelitian ini bertujuan untuk memperoleh konstruksi vektor ekspresi (pTarget) rekombinan yang mengandung sisipan gen hormon pertumbuhan ikan lele dumbo (Clarias sp.) dan promoter β-aktin ikan lele lokal (C. batrachus) dalam upaya pembuatan ikan lele lokal transgenik. Promoter β-aktin lele lokal (pCbBA) telah berhasil diisolasi dari hipofisa ikan tersebut dengan ukuran sekitar 1,7 kbp; dan memiliki elemen transkripsi CAAT box, TATA box, GC box, motif CarG, dan TGACG berdasar analisis program TF Bind. Penggantian promoter CMV (cytomegalovirus) yang terkandung dalam vektor ekspresi pTarget menggunakan dua enzim restriksi SgfI dan I-PpoI, menghasilkan fragmen DNA berukuran 6.083 bp (pTarget-GH lele dumbo) sebagai produk digesti. Fragmen pTarget-GH lele dumbo yang diligasi dengan promoter β-aktin lele lokal membentuk konstruksi By: Ibnu Dwi Buwono, Nono Carsono, Yuniar Mulyani, and M. Untung Kurnia

This study aims to obtain an expression vector construct (pTarget) containing recombinant growth hormone gene insertion of African catfish (Clarias sp.) and β-actin promoter derived from walking catfish (Clarias batrachus) in order to produce transgenic local catfish. β-actin promoter of walking catfish (pCbBA) have been isolated from the pituitary of the fish with a size of about 1.7 kbp, and has a transcription element: CAAT box, TATA box, GC box, CarG, and TGACG motif based on analysis result of TF Bind program. Replacement of CMV (cytomegalovirus) promoter contained in the expression vector pTarget using restriction enzymes SgfI and I-PpoI, obtained a product of digestion with the fragment size of 6,083 bp (pTarget-GH African catfish). pTarget-GH fragments were ligated with the African catfish β-actin promoter to arrange a construct of pTarget-pCbBA-African catfish GH (7,783 bp) as transgenic walking catfish expression vector.


Keywords


vektor ekspresi; hormon pertumbuhan; lele dumbo; lele lokal transgenik

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DOI: http://dx.doi.org/10.15578/jra.10.1.2015.11-24


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p-ISSN 1907-6754
e-ISSN 2502-6534


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