EFECTIVITY OF TRANSFER OF GROWTH HORMONE CCBA-CCGH GENE TO WILD BETTA, Betta imbellis EMBRYOS

The research was conducted to determine the successful introduction of genes encoding growth hormone (Growth hormone, GH) on Betta imbellis fish embryos.  GH gene of carp (CcGH) which is controlled by beta-actin promoter (CcBA) of carp as well.  GH gene of carp (CcGH) which is controlled by beta-actin promoter (CcBA) of carp also.  Betta imbellis embryo obtained from natural spawning the parents from Sumatra, collected 1-2 minutes after spawning at a density of 100 grains/2 mL.  Gene transfer was conducted by using transfection reagent X-treme Gene eplications.with different concentration 0,25 µL : 0,25 µL DNA GH gene dan 0,75 µL : 0,25 µL DNA GH gene; positif control (withot DNA plasmid) and negative control (without DNA plasmid and transfection reagent) with 5 replications.  Transfected embryos were hatched in palctic cans filled with 2 L water and placed in a water container.  The results of the experiment showed that the GH gene has been successfully transferred to the Betta imbellis embryo.  Hatching rate for all treatment about 80% and above.  Analysis of gene expression in seeds also showed activity GH gene expression in all treatments compared with positive and negative controls.  Total of individuals who carry the GH gene based analysis of fins per individual pieces as much as 16% and 13%.

Results of the analysis of genomic DNA in embryos Betta imbellis showed positive GH gene contained in all transfection treatment either in concentration of 0.25 mL and 0.75 mL. The success proves that the solution of the X-treme Gene transfection can be used as a medium for the introduction of foreign genes into fish embryos. Similar results were also reported by Prasetio et al (2012) that use a solution of X-treme Gene transfection has been used to insert genes fluorescent GFP (Green Flourenscent Protein). Use of the solution is not limited to Betta imbellis, but also can be used to fish the size of a small egg and spawn naturally. Embryos used for this study possible can amount greater than 200 embryos per container.

Analysis of transgene expression in embryonic fish Betta imbellis also showed activity characterized by the expression of genes in the DNA fragment targeted position. The research results show that the GH gene transfection, gene detection appear on treatment while the control (no transfection) tidk discovered fragments. Based on the description, for purposes of efficiency of use of 0.25 mL of solution as a material for gene transfer GH imbellis Betta fish is sufficient for use. As well as the use of a solution on the X-treme Gene in the manual issued is approximately 0.25 mL to 1 mL of the cell in general.