Cloning and Expression of ORF124 Koi Herpesvirus as a Vaccine

• Main Authors: Murwantoko ., Dewi Nur'aeni Setyowati, Rarastoeti Pratiwi, Masashi Kawaichi
• Format: Article
• Language: English
• Published: Universitas Gadjah Mada, Yogyakarta 2015-11-01
• Series: Indonesian Journal of Biotechnology
• Online Access: http://journal.ugm.ac.id/ijbiotech/article/view/7850

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table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin:0cm; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:11.0pt; font-family:"Calibri","sans-serif"; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:"Times New Roman"; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:"Times New Roman"; mso-bidi-theme-font:minor-bidi;} --> <!--[endif] --> <p class="MsoNormal"><em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua-Italic&quot;,&quot;serif&quot;;" lang="EN-GB">Koi herpesvirus </span></em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">(KHV) which also known as </span><em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua-Italic&quot;,&quot;serif&quot;;" lang="EN-GB">Cyprinid herpesvirus </span></em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">3 (CyHV-3), Koi herpes-like virus,</span></p> <p class="MsoNormal"><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">and carp interstitial nephritis gill necrosis virus (CNGV), caused signifi cant morbidity and mortality in koi</span></p> <p class="MsoNormal"><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">and common carp (</span><em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua-Italic&quot;,&quot;serif&quot;;" lang="EN-GB">Cyprinus carpio</span></em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">). The case fatality rate of this disease is 80&ndash;100%. Glycoprotein has been</span></p> <p class="MsoNormal"><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">used for vaccine development as sub unit vaccine against viruses. The aim of this research was to clone and</span></p> <p class="MsoNormal"><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">express membrane glycoprotein ORF124 KHV as a candidate of recombinant vaccine. ORF124 KHV gene was</span></p> <p class="MsoNormal"><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">successfully cloned into pBSKS and sequenced. Result showed that ORF124 KHV (isolate from Indonesia) had</span></p> <p class="MsoNormal"><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">100 % similarity with </span><em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua-Italic&quot;,&quot;serif&quot;;" lang="EN-GB">Cyprinid herpesvirus </span></em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">3 strain TUMST1 (from Japan), 99% similarity with </span><em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua-Italic&quot;,&quot;serif&quot;;" lang="EN-GB">Koi herpesvirus </span></em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">strain KHV-U (from USA) and </span><em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua-Italic&quot;,&quot;serif&quot;;" lang="EN-GB">Koi herpesvirus </span></em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">strain KHV-I (from Israel). Prediction analysis of T and B cell</span><em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua-Italic&quot;,&quot;serif&quot;;" lang="EN-GB"> </span></em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">epitopes showed that ORF124 KHV protein had 14 and 11 T cell epitopes (IAd, Rothbard/Taylor pattern),and had 10 B cell epitopes, suggested that the protein can be used as a vaccine candidate. ORF124 gene has</span><em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua-Italic&quot;,&quot;serif&quot;;" lang="EN-GB"> </span></em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">been expressed in </span><em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua-Italic&quot;,&quot;serif&quot;;" lang="EN-GB">Escherichia coli </span></em><span style="font-size: 9.5pt; font-family: &quot;BookAntiqua&quot;,&quot;serif&quot;;" lang="EN-GB">under pET32-a(+)vector.</span></p>