Fusion Strategy of Influenza A-H5N1 Virus M2e Epitope DNA Sequence to Hepatitis B Virus HBsAg-S Gene

DOI

10.2991/absr.k.200807.067How to use a DOI?

Keywords

A-H5N1 VLPs vaccine, asymmetric PCR, HBsAg-S, M2

Abstract

In order to make a broad vaccine for influenza-A virus, a conserve epitope must be considered. M2 is a conserve transmembran protein in all strain of influenza-A virus. Therefore, in this experiment we used 69 bp synthetic nucleotides from extracellular domain of M2 from H5N1 as an epitope (M2e) for influenza-A vaccine. A vaccine may usually more expose to immune system if they are made on a virus-like particles (VLPs). It was shown in many papers that the HBsAg-S protein is able to make a self-assemble VLP. Therefore, the synthetic M2e DNA was fused to a HBsAg-S gene, which encodes a small envelope protein of the Hepatitis B virus (HBsAg-S protein), which in turn creating a new VLP with M2e on it. This fusion replaced IPQ epitope of the HBsAg-S with M2e from M2 protein of the H5N1 virus. The fusion of M2e and HBsAg-S was conducted using asymmetric PCR. The single stranded DNA of M2e was produced with primers containing HBsAg-S gene sequence. Next, this single-stranded M2e was applied as mega primer for ‘fusion PCR’, which will fuse the M2e and part of the HBsAg-S fragment. A ~680 bp DNA band (fusion of M2e and HBsAg-S fragment) is generated from the ‘fusion PCR’. Alignment between hypothetic fusion sequence and the result was compared with native HBsAg-S sequence. The analysis shows that the IPQ epitope from HBsAg-S was totally replaced by the M2e.

Copyright

© 2020, the Authors. Published by Atlantis Press.

Open Access

This is an open access article distributed under the CC BY-NC license (http://creativecommons.org/licenses/by-nc/4.0/).

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TY  - CONF
AU  - Afifatul Achyar
AU  - Tati Kristianti
AU  - Sony Suhandono
PY  - 2020
DA  - 2020/08/07
TI  - Fusion Strategy of Influenza A-H5N1 Virus M2e Epitope DNA Sequence to Hepatitis B Virus HBsAg-S Gene
BT  - Proceedings of the International Conference on Biology, Sciences and Education (ICoBioSE 2019)
PB  - Atlantis Press
SP  - 333
EP  - 338
SN  - 2468-5747
UR  - https://doi.org/10.2991/absr.k.200807.067
DO  - 10.2991/absr.k.200807.067
ID  - Achyar2020
ER  -