| Summary: | Koi Herpesvirus (KHV) is one of the problems in carp and Koi (Cyprinus carpio) aquaculture. KHV can cause 100% mortality. Vaccination is one prevention method of viral diseases. Virus glycoprotein has been widely used as vaccine. The objectives of this research are to clone genes, express and purify proteins ORF25 KHV (encoding membrane glycoprotein) as a vaccine candidate. confirmed infected KHV of Carp from Magelang was used in this research. Primers were designed to amplify ORF25 by eliminating the hydrophobic
clusters. KHV�s ORF25 was successfully amplified and cloned in pET32a expression vector. Sequence analysis showed that Indonesian KHV�s ORF25 isolate has high simmilarity with cyprinid herpesvirus 3 DNA strain Japan, Israel, and America by 98%. T-cell epitope prediction using GENETYX, showed that
ORF25 has 9 iAd epitopes pattern and 8 Rothbard / Taylor pattern. B cell epitope prediction using "B Cell epitope Prediction Tools" from IEDB showed ORF25 has 3 epitops by Emini surface accessibility scale, 23 epitopes by Karplus and Schulz flexibility scale, 10 epitopes by Kolaskar and Tongaonkar antigenicity scale, 4
discontinue epitopes and 4 linear epitopes based Ellipro software. This result suggests that KHV�s ORF25 is predicted to be immunogenic. KHV ORF25 recombinant protein has been successfully produced. Production was achieved in E. coli BL21 cd (DE3). The protein was produced in insoluble form and optimal
when IPTG induction at OD 1, with a concentration of 1mM and incubated for 18 hours. Protein with a size of about 53 kd was successfully purified using Ni-NTA.
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