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Gene Cloning and Protein Expression of Koi Herpesvirus ORF25

HAYATI Journal of Biosciences · Vol. 23 No. 3 (2017)

DOI
10.4308/hjb.23.3.143
ISSN
2086-4094, 1978-3019
Terbit
2017-04-14
Penerbit
Bogor Agricultural University, Indonesia
Bahasa
eng

Abstrak

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.

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  • Diponegoro University
  • Universitas Gadjah Mada
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Rekan penulisOrganisasiArtikel bersama
Nur Lailatul Fitrotun NikmahUniversitas Gadjah Mada1
Wilis Ari SetyatiDiponegoro University1
SubagiyoDiponegoro University1
Muhammad ZainuddinDiponegoro University1
Erni MartaniUniversitas Gadjah Mada1

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Sumber metadata: OAI-PMH (metadata jurnal) http://journal.ipb.ac.id/index.php/hayati/oai, pertama di-harvest 8 Nov 2026, terakhir terlihat 8 Nov 2026.
Metadata asli (oai_dc)
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	<dc:title xml:lang="en">Gene Cloning and Protein Expression of Koi Herpesvirus ORF25</dc:title>
	<dc:creator xml:lang="en">Murwantoko, .</dc:creator>
	<dc:creator xml:lang="en">Fusiyanto, Cahya Kurnia</dc:creator>
	<dc:creator xml:lang="en">Triyanto, .</dc:creator>
	<dc:subject xml:lang="en">cloning</dc:subject>
	<dc:subject xml:lang="en">cyprinid herpesvirus 3</dc:subject>
	<dc:subject xml:lang="en">expression</dc:subject>
	<dc:subject xml:lang="en">ORF25</dc:subject>
	<dc:subject xml:lang="en">recombinant protein</dc:subject>
	<dc:subject xml:lang="en">vaccine</dc:subject>
	<dc:description xml:lang="en">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.</dc:description>
	<dc:publisher xml:lang="en">Bogor Agricultural University, Indonesia</dc:publisher>
	<dc:date>2017-04-14</dc:date>
	<dc:type>info:eu-repo/semantics/article</dc:type>
	<dc:type>info:eu-repo/semantics/publishedVersion</dc:type>
	<dc:type xml:lang="en">Peer-reviewed Article</dc:type>
	<dc:format>application/pdf</dc:format>
	<dc:identifier>http://journal.ipb.ac.id/hayati/article/view/16108</dc:identifier>
	<dc:identifier>10.4308/hjb.23.3.143</dc:identifier>
	<dc:source xml:lang="en">HAYATI Journal of Biosciences; Vol. 23 No. 3 (2016): July 2016; 143</dc:source>
	<dc:source>2086-4094</dc:source>
	<dc:source>1978-3019</dc:source>
	<dc:language>eng</dc:language>
	<dc:relation>http://journal.ipb.ac.id/hayati/article/view/16108/11902</dc:relation>
</oai_dc:dc>