Indonesian Aquaculture Journal
Vol 5, No 1 (2010): (June 2010)

OPTIMAL ELECTROPORATION CONDITION FOR SPERM MEDIATED GENE TRANSFER IN STRIPPED CATFISH (Pangasionodon hypophthalmus)

Raden Roro Sri Pudji Sinarni Dewi (Research Institute for Freshwater Fish Breeding and Aquaculture, Sukamandi)
Alimuddin Alimuddin (Department of Aquaculture, Faculty of Fisheries and Marine Science, Bogor Agricultural University, bogor)
Agus Oman Sudrajat (Department of Aquaculture, Faculty of Fisheries and Marine Science, Bogor Agricultural University, Bogor)
Komar Sumantadinata (Department of Aquaculture, Faculty of Fisheries and Marine Science, Bogor Agricultural University, Bogor)
Sularto Sularto (Research Institute for Freshwater Fish Breeding and Aquaculture, Sukamandi)



Article Info

Publish Date
30 Jun 2010

Abstract

The success of transgenic fish production has been achieved through eggs fertilization using electroporated sperms carrying exogenous DNA. This study was conducted in order to obtain the optimal electroporation condition for stripped catfish sperm. A plasmid containing green fluorescent protein (GFP) gene driven by carp β-actin promoter was transferred into sperm using electrophoresis method towards transgenic stripped catfish (Pangasionodon hypophthalmus) production. Electroporation was carried out using square wave shock with pulse length of 30 ms and pulse interval of 0.1 sec. Treatments are combination between voltage (50 V, 75 V, and 100 V) and pulse number (1 and 3). Exogenous DNA concentration used was 10 μg/mL of Tris-EDTA. Results showed that increasing the voltage from 50 to 100 decreased sperm motility, while pulse number did not affect sperm motility. Voltage of 50 gave the best motility of sperm, although sperm viability relatively similar between treatments and control except at 100 V with 3 pulses number. Further, electroporation-treated sperms were able to fertilize eggs. Higher hatching rate of eggs was obtained in electroporation treatment at 50 V with pulse number of 1 and 3. The persistence of transferred GFP was detected in electroporated and incubated sperms (control). However, GFP was only detected in larvae from eggs that were fertilized by electroporated sperm. Thus, electroporation could be applied to produce transgenic stripped catfish. 

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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 ...