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.
Penulis: Slamet Subyakto, Gemi
Triastutik, Rustidja, Komar Sumantadinata, Alimuddin, M. Sasmito Jati, Irvan
Faizal, Ratu Siti Aliah
Kode Jurnal: jpperikanandd100163
