Indonesian Aquaculture Journal
Vol 11, No 1 (2016): (June 2016)

GENE TRANSFER ON Betta imbellis THROUGH TRANSFECTION METHOD WITH DIFFERENT DNA CONCENTRATION

Eni Kusrini (Research and Development Institut for Ornamental Fish Culture Jl. Perikanan No.13 Pancoran Mas Depok 16436)
Alimuddin Alimuddin (Aquaculture Post-graduate program, Department of Aquaculture,FPIK,IPB Department of Aquaculture, Marine and Fishery Faculty, Bogor Agricultural University Jl. Rasamala, Kampus IPB Dramaga, Bogor 16680.)
Mohammad Zairin (Aquaculture Post-graduate program, Department of Aquaculture,FPIK,IPB Department of Aquaculture, Marine and Fishery Faculty, Bogor Agricultural University Jl. Rasamala, Kampus IPB Dramaga, Bogor 16680.)
Dinar Tri Sulistyowati (Aquaculture Post-graduate program, Department of Aquaculture,FPIK,IPB Department of Aquaculture, Marine and Fishery Faculty, Bogor Agricultural University Jl. Rasamala, Kampus IPB Dramaga, Bogor 16680.)



Article Info

Publish Date
28 Dec 2016

Abstract

Big size betta (Giant) have a high economic value compared to normal size betta, and over expression of growth hormone gene can produce giant fish.  As an initial step of giant transgenic betta productions, this study was conducted in order to obtain DNA plasmid concentration which provide higher hatching and survival rate of betta larvae.  Construction of PhGH pCcBA gene contains growth hormone gene of Siamese catfish (PhGH) and it is controlled by the CCBA promoter. Betta imbellis broodstocks were spawned naturally, and embryos were collected 1-2 minutes after spawning time. One hundred embryos were dipped in 2 mL of transfectan X-treme gene which containp CcBA-PhGH construction genes (50 µg/mL), on room temperature for about 30 minutes. Treatments on this study were different transfectant : DNA plasmid ratiosnamely:A (0,75 µL: 0,25 µL); B (0,75 µL : 0,50 µL); C (0,75 µL: 0,75 µL), D as Control 1(without transfectant, 0,25 µL DNA); E.as Control 2(0,75 µL transfectant, without DNA), and Fas control 3 (without transfectant and without DNA). Every treatments was repeated three times.  Transfection embryos were hatched on a container (1L Volume). Study results showed that hatching rate and larvae survival rate  (4 days after hatching) on treatment A were the same with the control, but slightly higher than B and C treatments. PCR analysis with DNA template showing that PhGH gene were found on embryos and larvae (pooled sample) of treatment A, B and C. Furthermore, RT-PCR analysis showing the existence of mRNA PhGH expression on embryos and larvae (pooled sample). Therefore, embryo transfection with transfectant ratio 0,75 µL and  DNA 0,25 µLshowing the best results.

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Journal Info

Abbrev

iaj

Publisher

Subject

Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology

Description

Indonesian Aquaculture Journal is a peer-reviewed and open access journal based in Indonesia that globally/internationally accepts and publishes scientific articles in the field of aquaculture. The journal is hosted and managed by the Center for Fisheries Research, Indonesian Ministry of Marine ...