Gene Cloning and Protein Expression of Koi Herpesvirus ORF25

Koi herpesvirus (KHV) caused significant morbidity and mortality in koi and common carp (Cyprinus carpio). Glycoprotein has been used for vaccine development as sub unit vaccine against many viruses. KHV ORF25 is one of koi herpesvirus genes which encode a glycoprotein. The objectives of this resear...

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Main Authors: Murwantoko, (Author), Fusiyanto, Cahya Kurnia (Author), Triyanto, (Author)
Format: EJournal Article
Published: Bogor Agricultural University, Indonesia, 2017-04-14.
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LEADER 02429 am a22003133u 4500
001 HAYATI_16108_11902
042 |a dc 
100 1 0 |a Murwantoko, .  |e author 
700 1 0 |a Fusiyanto, Cahya Kurnia  |e author 
700 1 0 |a Triyanto, .  |e author 
245 0 0 |a Gene Cloning and Protein Expression of Koi Herpesvirus ORF25 
260 |b Bogor Agricultural University, Indonesia,   |c 2017-04-14. 
500 |a https://journal.ipb.ac.id/index.php/hayati/article/view/16108 
520 |a Koi herpesvirus (KHV) caused significant morbidity and mortality in koi and common carp (Cyprinus carpio). Glycoprotein has been used for vaccine development as sub unit vaccine against many viruses. KHV ORF25 is one of koi herpesvirus genes which encode a glycoprotein. The objectives of this research are to clone gene KHV ORF25and express its protein. The common carp showing necrosis and white patches of gill which was collected from Magelang was used in this research. Primers were designed to amplify partial ORF25 based on KHV J strain. KHV ORF25 was successfully amplified and cloned in pET32a. Sequence analysis showed that this KHV ORF25 has 99% homology with the sequences of KHV genotype KHV-J, KHV-I, and KHV-U. This ORF was predicted has 3, 23, and 8 B-cell epitopes based on Emini scale, Karplus and Schulz scale, and ElliPro respectively. The KHV ORF25 recombinant protein has been successfully produced in Escherichia coli as an insoluble protein with approximately 45 kDa in size. The high protein production was achieved when the protein induction was done at bacterial density at OD600 as 1.0 with 1-mM isopropyl β-D-1-thiogalactopyranoside (IPTG) and incubated at 37°C for 18 hours. The protein predicted has immunogenicity and the potency as a vaccine is needed to be evaluated. 
546 |a eng 
690 |a cloning 
690 |a cyprinid herpesvirus 3 
690 |a expression 
690 |a ORF25 
690 |a recombinant protein 
690 |a vaccine 
655 7 |a info:eu-repo/semantics/article  |2 local 
655 7 |a info:eu-repo/semantics/publishedVersion  |2 local 
655 7 |a Peer-reviewed Article  |2 local 
786 0 |n HAYATI Journal of Biosciences; Vol. 23 No. 3 (2016): July 2016; 143 
786 0 |n 2086-4094 
786 0 |n 1978-3019 
787 0 |n https://journal.ipb.ac.id/index.php/hayati/article/view/16108/11902 
856 4 1 |u https://journal.ipb.ac.id/index.php/hayati/article/view/16108/11902  |z Get fulltext