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OJVRTM
Online Journal of Veterinary Research©
Volume 17 (1): 28-33, 2013.
Extensively Redacted 2017.


Expression of constant region of bovine IgG1 heavy chain in E. coli

 

Abdolhossein Jangeran Nejad1, Atefeh Ashtary1, Masoud Ghorbanpoor2 and Masoudreza Seyfi Abad Shapouri2

 

1DVM, 2DVM, PhD, Department of Pathobiology, School of Veterinary Medicine, Shahid Chamran University, Ahvaz, Iran

 

ABSTRACT

 

Nejad AJ, Ashtary A, Ghorbanpoor M, Seyfi M, Shapouri A., Expression of the constant region of bovine IgG1 heavy chain in E. coli, Online J Vet Res., 17 (1): 28-33, 2013. We describe expression of constant region of bovine IgG1 heavy chain in E. coli for antibody production. Specific primers were used to amplify constant region of G1 chain mRNA by RT-PCR. The resulting DNA fragment was cloned in pMal-C2X expression plasmid and expressed in TG1 and BL strains of E. coli. Expression of an expected fusion protein (MBP-G1), comprising Maltose binding protein (MBP), encoded by the plasmid and the constant region of G1 chain (CH1-CH3) was confirmed by SDS-PAGE and Western blot. The protein was found to be insoluble in BL21(DE3) strain but partially soluble in TG1, when expressed at room temperature. Our results suggest that the constant region of bovine IgG1 heavy chain can be expressed as a partially soluble fusion protein in E. coli. This strategy is simple and when established, eliminates the time consuming steps of immunoglobulin purification. Moreover, by this strategy it is possible to express the isotype specific sequences of immunoglobulins and facilitate the production of isotype specific anti antibodies.

 

 

Key Words: Bovine, IgG1, constant region, expression, E. coli


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