', Alimuddin
Lembaga Penelitian dan Pengabdian kepada Masyarakat Universitas Riau

Published : 2 Documents
Articles

Found 2 Documents
Search

Aktivitas Promoter â-aktin Ikan Medaka Jepang (Oryzias latipes) pada Ikan Mas (Cyprinus carpio)

Jurnal Natur Indonesia Vol 11, No 02 (2009)
Publisher : Lembaga Penelitian dan Pengabdian kepada Masyarakat Universitas Riau

Show Abstract | Original Source | Check in Google Scholar | Full PDF (72.918 KB)

Abstract

This study was conducted to examine activity of medaka (Oryzias latipes) â-actin promoter (mBP) in common carp(Cyprinus carpio) as the first step towards development of common carp transgenic in country. Gene constructpmBP-hrGFP that consists of mBA promoter and humanized Renilla reniformis green fluorescent protein gene(hrGFP) was injected into cytoplasm of one cell stage of common carp by using microinjector. PmBP-hrGFPconcentration used for microinjection was 50 μg/mL aquabides. Parameters observed were survival rate of embryo(SRe), hatching rate (HR) and expression of hrGFP gene. SRe was calculated before eggs hacthed, while hatchingrate (HR) was after all of eggs hatched. The activity of mBA promoter was analyzed by observation of hrGFP genetransient expression using a fluorescence microscope. The results of experiment showed that SRe (87,5%) andHR (79.2%) of control was respectevily higher than that of injected treatment (75.0% & 61.7%). Expression of hrGFPwas observed firstly at blastula (12 hours after fertilization) to 1-day-old larval stages (24 hours after hatching)with higher gene expression at blastula to late gastrula stages. Percentage of micronjected larvae expressinghrGFP at 6 hours after hatching reached 71.6 ± 6.7%. Conclusion was that mBA promoter could drove hrGFPexpression in common carp, hence it can be used to produce common carp transgenic by changing hrGFP withgenes correlated with important traits in aquaculture.

Construction of a DNA Vaccine Using Glycoprotein Gene and Its Expression Towards Increasing Survival Rate of KHV-Infected Common Carp (Cyprinus carpio)

Jurnal Natur Indonesia Vol 13, No 01 (2010)
Publisher : Lembaga Penelitian dan Pengabdian kepada Masyarakat Universitas Riau

Show Abstract | Original Source | Check in Google Scholar | Full PDF (120.942 KB)

Abstract

Deoxyribonucleic acid (DNA) vaccine has recently been developed as an alternative vaccine against virus infection.This study was the first step of DNA vaccine development to protect cyprinids including common carp (Cyprinuscarpio) and fancy koi (Cyprinus carpio) from KHV (koi herpesvirus) infection in Indonesia. One of KHV glycoproteingenes, i.e. glycoprotein (GP) was ligated with Japanese medaka (Oryzias latipes) â-actin promoter to generatepAct/GP as a DNA vaccine. Fourty fish in body weight of 10-15 g/fish were individually injected by pAct/GP intomuscle in different dosage of 2.5 μg, 7.5 μg and 12.5 μg/100 μl phosphate buffer saline. Total RNA was extractedfrom the 12.5 μg of pAct/GP-injected fish muscle at 24, 48 and 67 hours post-injection to analyze GP expression byRT-PCR method. Potential of pAct/GP as DNA vaccine was examined by injecting KHV into the 30-days-vaccinatedfish. Both of possitive and negative control fish group were not vaccinated. Possitive control fish group wereinjected with KHV, but negative control fish group were not. KHV-challenged fish were reared for 1 month, and thedeath fish were calculated daily. Result of RT-PCR analysis showed that GP gene expression were detected at 3 dpost-injection. Expression of GP in the vaccinated fish groups helped to improve their survival rate after challengedby KHV. All of fish without DNA vaccination had dead 17 days after KHV injection. The results demonstrated thatpAct/GP had high potency to be used as a DNA vaccine against KHV infection in cyprinids.