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

Slamet Subyakto¹, Gemi Triastutik¹, Rustidja², Komar Sumantadinata³, Alimuddin³, M. Sasmito Jati⁴, Irvan Faizal⁵, Ratu Siti Aliah⁵

1. Balai Budidaya Air Payau Situbondo, Direktorat Jenderal Perikanan Budidaya, DKP. E-mail: slamet_subyakto@yahoo.com
2. Fakultas Perikanan dan Kelautan, Universitas Brawijaya, Malang
3. Departemen Budidaya Perikanan Fakultas Perikanan dan Kelautan, IPB
4. Fakultas Matematika dan Ilmu Pengetahuan Alam, Universitas Brawijaya, Malang
5. Badan Pengkajian dan Penerapan Teknologi, Kementrian Negara Riset dan Teknologi, Jakarta

Diterima 5 April 2010; disetujui 10 Mei 2010

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

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