Murwantoko, Fusianto, Cahya Kurnia, and Triyanto (2016) Gene Cloning and Protein Expression of Koi Herpesvirus ORF25. HAYATI Journal of Biosciences, 23. pp. 143-149.

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Abstract

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 ORF25 and 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.

Item ID:	90899
Item Type:	Article (Research - C1)
ISSN:	2086-4094
Keywords:	ORF25,cloning,cyprinid herpesvirus 3,expression,recombinant protein,vaccine
Copyright Information:	Copyright © 2016 Institut Pertanian Bogor. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
Date Deposited:	24 Sep 2026 05:29
FoR Codes:	30 AGRICULTURAL, VETERINARY AND FOOD SCIENCES > 3005 Fisheries sciences > 300503 Fish pests and diseases @ 40% 30 AGRICULTURAL, VETERINARY AND FOOD SCIENCES > 3001 Agricultural biotechnology > 300103 Agricultural molecular engineering of nucleic acids and proteins @ 60%
SEO Codes:	10 ANIMAL PRODUCTION AND ANIMAL PRIMARY PRODUCTS > 1002 Fisheries - aquaculture > 100202 Aquaculture fin fish (excl. tuna) @ 100%
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