©1996-2015 All Rights Reserved.
Online Journal of Veterinary Research . You may not store these pages in any
form except for your own personal use. All other usage or distribution is
illegal under international copyright treaties. Permission to use any of these
pages in any other way besides the before mentioned must be gained in writing
from the publisher. This article is exclusively copyrighted in its entirety to
OJVR. This article may be copied once but may not be, reproduced or
re-transmitted without the express permission of the editors. This journal
satisfies the refereeing requirements (DEST) for the Higher Education Research
Data Collection. Linking:To link to this page or any pages linking to this page
you must link directly to this page only here rather than put up your own page.
OJVRTM
Online Journal of Veterinary Research©
Volume 19(2): 124-129, 2015.
Cloning, expression and purification of M2e-HA2 from
Influenza A virus in Escherichia coli
Masoud Moghadaszadeh1,*,
Mehdi Golchin2, Hadi Tavakkoli3,
Reza Ghanbarpour2
12Department(s) of
Pathobiology, 3Clinical Sciences, School of Veterinary Medicine, Shahid Bahonar University of
Kerman, Kerman, Iran.
ABSTRACT
Moghadaszadeh M, Golchin M, Tavakkoli H, Ghanbarpour R., Cloning, expression and purification of
M2e-HA2 from Influenza A virus in Escherichia
coli, Onl J Vet Res., 19(2): 124-129, 2015. Influenza can be controlled by
vaccination but the virus due to its high mutation rate, has limited efficacy. Influenza
viruses possess conserved epitopes matrix-2 protein ecto-domain
(M2e) and HA2 that could be used in a vaccine. In this work, M2e-HA2 peptide was
expressed in E. coli and then purified. The M2e-HA2 gene from influenza A virus was then synthesized and cloned into pGS21vector.
The construct was transferred into E. coli BL21 (DE3) and induced using
IPTG. Expression of recombinant peptide was confirmed by western blot assay
using anti-GST monoclonal antibody. The expressed peptide-GST was purified from
bacterial lysate by IMAC chromatography. The resulting sequence revealed that
the M2e-HA2 gene was cloned in the vector. SDS-PAGE electrophoresis of purified
peptide demonstrated a strong single band. Western blot analysis showed a
single band in correct position. The purified peptide could be tested In vivo against influenza virus
infection.
Key words: Cloning,
Expression, Purification, M2e-HA2, Escherichia coli, Influenza vaccine.
FULL-TEXT(SUBSCRIBERS) OR PURCHASE ARTICLE