The Effectiveness of hrGFP Gene Reporter Role in Carp Fish (Cyprinus carpio) Transgenesis Process Based on Convocal Microscopy Analysis

Abstract

Sperm cells can be used as a medium to insert foreign DNA into the egg. Electroporation humanized renilla reformis Green Fluorescent Protein (hrGFP) gene as a reporter gene is very important to know the effectiveness of gene transfer. Research is aimed to understand the effectiveness rate of hrGFP gene reporter to detect the successful gene transfer process based on the result of observation with convocal microscope over transgenic sperm level, embryo and larva of Carp Fish. Result of trans-genesis of hrGFP as sperm-mediated gene reporter must be accurately detected. Based on the result of observation with convocal microscope, it is shown that hrGFP enters the core level, indicates by PI pigmentation and thereby, hrGFP gene reporter can provide information about hrGFP fluorescence starting from sperm, embryo and larva of Carp Fish.

The Effect of Electroporation Method towards the Motility and Viability of Java Barb Fish (Puntius javanicus) Sperm

“…This is different from the results of Japanese Carp (Cyprinus carpio var.Koi) Luthfiyah et al [15] which obtained the best results on 10 V treatment with motility score 4 and the lowest at 40 V with motility a score of 0.5 [15]. The treatment in this study without using level voltage (10V) because of based on the result from previous research in carp fish (Cyprinus carpio) by electroporation method on sperm with the best result treatment was voltage level (30V) [9]. Further, the motility and viability of Java barb fish sperm using treatment with voltages level among…”

“…Determination treatment with different voltages level (20 V, 30 V, 40 V) in this study based on the result of previous research on Japanese carp (Cyprinus carpio var.Koi) sperm using electroporation method with voltage level 30 V [9]. In each of the treatment, the electric shock was given for 1 ms repeated 4 times, except for the control [9].…”

“…In each of the treatment, the electric shock was given for 1 ms repeated 4 times, except for the control [9].…”

“…After that, the voltage of square wave was chosen and adjustment on the strength of the electricity, shock duration and the shock frequency were made before the pulse button was pushed to start the shock. This treatment was repeated 3 times for each of the voltage [9].…”

“…After theat, the sperm was poured into the appendorf and each of the 25 µl of sperm was observed to see its motility and its viability [9].…”