PENGGUNAAN METODE MIKROINJEKSI DAN ELEKTROPORASI DALAM TRANSFER GEN HORMON PERTUMBUHAN (GH) PADA IKAN KERAPU TIKUS (Cromileptes altivelis)

ABSTRACT: Humpback grouper is one of the mariculture fish species that possess high economical price in Asia, including Indonesia. The main problem facing in humpback grouper culture is its low growth rate. Transgenesis technique may be one of the methods to solve the problem. This study was conducted to determine the effectiveness of microinjection and electroporation techniques to transfer of growth hormone gene in humpback grouper. Microinjection was carried out to 1-4 cell stage embryos using 25 µg/ml of foreign DNA solution, and duration of injection into each embryo was 1, 2 and 3 seconds. Electroporation by 50 V, 30 ms of pulse length, 5 of pulse number and 0.1 of pulse interval was performed to sperm using three DNA concentration of 5, 10 and 20 µg/ml. The incorporation of foreign DNA in sperm and embryos were analyzed using PCR method. Based on PCR analysis, an optimum DNA concentration for electroporation was 10 µg/ml. Limited number of embryos could be microinjected during 20-30 min of time to reach 4 cells stage embryos. Microinjection for 1 second was allowed higher number of embryos survived, although no or very low number of larvae was hatched. Our study revealed that microinjection as well as electroporation could be used as transgenesis methods for humpback grouper. By means of simplicity and efficacy, however, electroporation was an appropriate gene transfer method for humpback grouper.
Keywords: Microinjection, electroporation, transgenesis, growth hormone, humpback grouper
Penulis: Slamet Subyakto, Gemi Triastutik, Rustidja, Komar Sumantadinata, Alimuddin, M. Sasmito Jati, Irvan Faizal, Ratu Siti Aliah
Kode Jurnal: jpperikanandd100163

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