Kloning dan Sekuensing Gen Penyandi Protein Permukaan VP19 WSSV Isolat Situbondo

Abstract

WSSV (White Spot Syndrome Virus) is the most dangerous shrimp pathogen and give a very bad impact to the shrimp industry in many countries, including Indonesia. Recombinant vaccine of viral protein-19 (VP19) from abroad is able to increase the relative survival rate of P. monodon about 77% when challenged with WSSV. However, this vaccine is still difficult to obtain, expensive, limited availability, and unsustainable. Furthermore, the vaccine usage is less effective since the local WSSV suspected to have some mutation and have differences in sequence from the one that used for recombinant vaccine production so far. Therefore, it is necessary to produce recombinant vaccine for immunization of local shrimp according to WSSV isolates from Indonesia. The objectives of this study are to obtain VP19-encoding gene of WSSV from Situbondo isolates in order to overcome WSSV infection and know the moleculer technology related with cloning certain encoding protein gene of WSS virus causing desease that generally attacks shrimps. This research conducted through several stages DNA genome isolation of P. monodon infected with WSSV, amplification of VP19-encoding gene using PCR, isolation of VP19-encoding gene from agarose gel, ligation of VP19- encoding gene into pGEM-T Easy vector, pGEM-T Easy vector transformation to E.Coli, recombinant vector screening, vector isolation using mini preparation technique, vector verification and sequencing. Cloning and Sequencing of VP19-encoding Gene of WSSV From Situbondo isolates has been conducted. The result of cloning shows that the size of VP19-encoding gene from Situbondo isolates is similar to VP19-encoding gene from the abroad one, which is 366 bp. However, the sequence result shows that there are some differences on bases and amino acid. However the result of sequencing shows there are difference of 5 base and 3 amino acid arranged VP19 sequence the Situbondo isolate with isolate from several countries. The similiarity the Situbondo isolate sequence with isolate from Indonesian itself altough from other countries is 98%.