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

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.

Pengaruh Padat Tebar terhadap Konsumsi Oksigen dan Respons Stres Ikan Cupang Alam (Betta imbellis)

“…Oleh karena itu, budidaya ikan ini sangat mudah karena tidak membutuhkan peralatan untuk meningkatkan kadar oksigen terlarut pada media pemeliharaannya. Beberapa penelitian pada ikan cupang alam ini juga sudah dilakukan, di antaranya yaitu terkait pemijahan dan pemeliharaan larva (Murniasih et al, 2012), perkembangan embrio (Cindelaras et al, 2015), pertumbuhan larva pada suhu pemeliharaan yang berbeda (Rahmawati & Kusrini, 2016), performa pertumbuhan dengan pemberian hormon (Hayuningtyas & Kusrini, 2016), transfer gen (Prasetio et al, 2013;Kusrini et al, 2016), dan performa hasil transgeniknya (Kusrini et al, 2018).…”

Crossing and gene flow of the transgens of the red fluorescent proteins (RFP) and green (GFP) in the zebrafish (Danio rerio) transgenic introduced to Peru

“…Sin embargo, existe la necesidad de implementar y estándarizar protocolos moleculares para la detección de la presencia y flujo génico de los transgenes de las proteínas de fluorescencia en OVM´s como son los peces Cebra (Danio rerio) transgénicos ornamentales introducidos al país y actualmente existentes en varios acuarios o criaderos comerciales donde son criados y reproducidos libremente en el Perú. Mundialmente, existe una creciente preocupación ante la falta de información generada por una valoración real de los efectos negativos por la introducción, reproducción e hibridación descontrolada (Flujo génico de un organismo transgénico a un organismo no transgénico emparentado) de éstos peces modificados genéticamente en sus colores corporales u otros rasgos por la acción de transgenes de organismos marinos invertebrados (Kusrini et al, 2016;Mohanta et al, 2014;Ofelio et al, 2012;Scotto, 2016).…”

Gene Transmission, Growth, and Exogeneous Growth Hormone Expression of G2 Transgenic Betta Fish (Betta imbellis)

“…The fish were maintained in the controlled indoor rearing facility in the Research Institute for Ornamental Fish Aquaculture, Depok, West Java, Indonesia. Previously, The G 0 transgenic fish showed 37.8% higher body weight than non-transgenic control after 7 months of rearing, and the PhGH cDNA was detected in the various tissues after PCR amplification (Kusrini et al, 2016a;Kusrini et al, 2016b). Furthermore, the G 1 transgenic was produced from three different crosses of G 0 : transgenic (TG) male and non-transgenic (NT) female, NT male paired with TG female, and NT transgenic fish pair as a control.…”

“…In our recent study, we have successfully produced G 0 and G 1 GH-transgenic fish of the B. imbellis, native ornamental betta from Indonesia (Kusrini et al, 2018;Kusrini et al, 2016a). The gene introduction was conducted using the pCcBA-PhGH vector that consisted of the P. hypophthalmus GH cDNA and regulated by C. carpio β-actin promoter (Dewi et al, 2012).…”