KLONING DAN EKSPRESI GEN ORF124 KOI HERPESVIRUS SEBAGAI KANDIDAT VAKSIN

Dewi Nuraeni Setyowati, S.Pi., Dr. Ir. Murwantoko, M.Si.,

2012 | Tesis | S2 Bioteknologi

Koi herpesvirus (KHV) yang dikenal juga sebagai Cyprinid herpesvirus 3 (Cy-HV 3), Koi herpes-like virus, dan carp interstitial nephritis gill necrosis virus (CNGV) menyebabkan penyakit dan kematian pada ikan koi dan karper (Cyprinus carpio). Kematian karena penyakit KHV mencapai 80-100% pada populasi yang terinfeksi. Glikoprotein telah banyak digunakan untuk pengembangan vaksin sub unit. Tujuan dari penelitian adalah mengklon dan mengekspresikan gen ORF124 KHV penyandi glikoprotein membran sebagai kandidat vaksin rekombinan. Gen ORF124 KHV telah berhasil diklon ke pBSKS dan disekuensing. Hasil penelitian menunjukkan bahwa ORF 124 KHV isolat Indonesia mempunyai kesamaan dengan Cyprinid herpesvirus 3 DNA strain TUMST1 (AP008984.1) isolat Jepang sebesar 100%, mempunyai kesamaan dengan Koi herpesvirus strain KHV-U (DQ657948.1) isolat Amerika dan Koi herpesvirus strain KHV-I (DQ177346.1) isolat Israel sebesar 99%. Analisis prediksi epitop sel T dan sel B menunjukkan bahwa protein ORF124 dapat digunakan sebagai kandidat vaksin karena memiliki 14 dan 11 epitop sel T (pola IAd dan Rothbard/Taylor) serta memiliki 10 epitop sel B. Gen ORF124 KHV berhasil diekspresikan menggunakan vektor pET32- a(+).

Koi herpesvirus (KHV) which also known as Cyprinid herpesvirus 3 (CyHV-3), Koi herpes-like virus, and carp interstitial nephritis gill necrosis virus (CNGV), caused significant morbidity (sickness or disease) and mortality in koi and common carp (Cyprinus carpio). Mortality of KHV disease reached 80–100% in affected populations. Glycoprotein has been used for vaccine development as sub unit vaccine. The aim of this research was to clone and express membrane glycoprotein ORF124 KHV as a candidate of recombinant vaccine. ORF124 KHV gene was successfully cloned into pBSKS and sequenced. Result showed that ORF124 KHV (isolate from Indonesian) had 100 % similarity with Cyprinid herpesvirus 3 DNA strain TUMST1 (AP008984.1, isolate from Japan), 99% similarity with Koi herpesvirus strain KHV-U (DQ657948.1, isolate from USA) and Koi herpesvirus strain KHV-I (DQ177346.1, isolate from Israel). Prediction analysis of T and B cell epitopes showed that ORF124 KHV protein can be used as a vaccine candidate because had 14 and 11 T cell epitopes (IAd, Rothbard/Taylor pattern), and had 10 B cell epitopes. ORF124 gene can be expressed in pET32-a(+).

Kata Kunci : Koi herpesvirus, ORF124, vaksin, glikoprotein