UNS Conference Portal, The 1st International Conference on Science, Mathematics, Environment and Education 2017

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Cloning and Molecular Analysis of Hepatitis C Virus (HCV) 1a NS4b Isolated in Central Java, Indonesia
Faqihuddin Ahmad, Afiono Agung Prasetyo, Yulia Sari, Risya Cilmiaty Arief Riswiyant

Last modified: 2017-07-16


Hepatitis C virus (HCV) nonstructural protein 4B (NS4B) is a multi-transmembrane protein, essential for HCV replication and pathogenesis; however, the contribution of NS4B is still little known. To understanding the potential role of HCV NS4B, a complete coding sequence of HCV 1a NS4B was cloned and molecular analyzed. RNA HCV was isolated from an anti HCV positive plasma sample in a molecular epidemiology study in Central Java, Indonesia, followed by cDNA synthesis and RNA HCV molecular detection by a nested PCR, respectively. NS4B gene was then amplified and inserted into an Escherichia coli expression plasmid. The recombinant plasmid was transformed into competent cells, extracted, sequenced and genotyped. The sequence result was aligned and analyzed with all of HCV sequences with same genotype in GenBank, and then molecular analyzed. The HCV genome isolated was genotyped as HCV 1a. In HCV NS4B molecular analysis, the NS4B sequence of HCV isolate was consistent with that of previous reports. However, amino acid variations were found in N terminal domain, transmembrane domain, and C terminal domain. The H2 sequence at C terminal domain and palmitoylation site motif consensus were also proposed.