Introduction and Expression of Growth Hormone Gene on Transgenic Common Carp

Abstract

Common carp Cyprinus carpio is one of the economically valuable aquaculture fish species in Indonesia. This research was aimed to produce transgenic common carp that grow rapidly by over-expressing the tilapia growth hormone (tiGH) gene. Trangenic common carp were produced by sperm-mediated gene transfer using electroporation method. In this study, two electroporation programs that developed by other researcher were applied to obtain a suitable electroporation method for common carp. The electroporated sperm were then used to fertilize eggs to produce F0 transgenic fish. Transgenic F0 carrying tiGH in their sperm were crossed with non-transgenic fish to produce transgenic first (F1) generation. Identification of transgenic fish was performed by PCR method using specific primer for tiGH gene. Expression of tiGH was analyzed using RT-PCR method and growth measurement at F1 fish generation. The results showed that electroporation of sperm using 50 Volt/cm, and DNA concentration of 25 ng/μl was produced higher percentarge of fish carrying tiGH in their gamet compared to that of another electroporation program (40 Volt/cm, DNA concentration of 50 ng/μl). Five sexually matured males F0 transgenic fish were selected and mated with non-transgenic to produce F1 generation. Percentage of F1 progenies carrying tiGH was 47,2-66,7%. Variated measure of F1 genereations was 10,1->12,0 cm size (22,5%-42,9%), 8,1->10,0 cm size (14,3%-42,9%), 6,1->8,0 cm size (11,9%-29,2%), 4,1->6,0 cm size (0,0%-39,3%). Increasing of growth of F1 was about 1,4-1,87 times for non transgenic fish. Survival of transgenic and non-transgenic F1 was generally similar. RT-PCR analysis showed that the F1 transgenic fish have tiGH gene expression in caudal fin, while no expression in non-transgenic fish were found. Thus, the increased body weight of F1 transgenic common carp was associated with expression of tiGH gene, and this fish can be used as broodstock for the production of fast-grow stable transgenic common carp.