Title of article :
Fusion of CtxB with StxB, Cloning and Expression of in Esherichia coli: A challenge for Improvement of Immune Response Against StxB
Author/Authors :
Madanchi, Hamid Department of Biology - Imam Houssein University, Tehran , Honari, Hossein Department of Biology - Imam Houssein University, Tehran , Sadraeian, Mohammad Department of Biology - Imam Houssein University, Tehran , Hesaraki, Mahdi Department of Biology - Imam Houssein University, Tehran
Abstract :
Cholera toxin B subunit (CtxB) is a homopantameric, nontoxic subunit of
cholera toxin that is responsible for its binding to the cell and has been known as
a mucosal adjuvant for vaccines that could increase homoral and mocusal immunity
response. In this work, the CtxB gene was fused to the StxB gene from Shigella
dysenteriae type I a vaccine antigen candidate against this pathogen, by a nonfurin
linker then ligated with pGEM vector and subcloned in the pET28a(+) as an
expression vector . The CtxB-StxB fusion protein was expressed in Escherichia coli,
and purified by a Ni-NTA resin column, then detected molecular weight and
immunogenicity by SDS-PAGE and Western-blot. StxB has low molecular weight,
so immune response against it is low, while CtxB is a potent mucosal adjuvant. In
this method, the CtxB-StxB fusion protein was expressed in Escherichia coli in order
to use its natural adjuvanticity of the CtxB which will enhance immune response
against StxB, as well as, it will produce immune response against both shiga and
cholera toxins.
Keywords :
Adjuvant , Expression , CtxB , Shigella toxin B subunit (StxB) , Shigella dysenteriae
Journal title :
Astroparticle Physics