Expression and purification of HCMV glycoprotein B full protein in Escherichia coli

Somayeh Azadfar, Ahad Yamchi, Ahmad Majd, Alijan Tabarraei

2020, Vol: 11, Issue: 3

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J Complement Med Res. 2020; 11(3): 60-67

doi: 10.5455/jcmr.2020.11.03.08

Expression and purification of HCMV glycoprotein B full protein in Escherichia coli

Somayeh Azadfar, Ahad Yamchi, Ahmad Majd, Alijan Tabarraei.

Abstract
Background: Human cytomegalovirus (HCMV) is associated with significant mortality in immunocompromised individuals and neonates and is a major concern of global public health. Some viral glycoproteins such as glycoproteins B have an important role on virus life. This herpesvirus fusion protein is a candidate for the vaccine due to its undeniable role in infection.
Materials and Methods: Here we designed a full-length gB sequence optimization to increase the expression level of this protein. Numerous factors such as the adaptation index of codon, codon context, and adapted GC content based on E. coli codon usage. In addition, the ribosome binding site (RBS) of pET-15b was redesigned. Various factors such as IPTG concentration and induction time were optimized. The recombinant gB protein was purified by the Ni-sepharose column.
Result: Among IPTG concentrations, and post inductions, the level of gB was higher in 1mM IPTG, and 8 hours, respectively. In hybrid purification conditions, gB protein was purified completely.
Conclusion: The protein designed in this study be expressed in the prokaryotic system and with an optimal design, it can be purified properly. Our data showed that it can be used as an effective vaccine in vivo immunogenicity evaluation in the future.

Key words: cytomegalovirus, glycoprotein, protein purification, vaccine

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Pubmed Style

Somayeh Azadfar, Ahad Yamchi, Ahmad Majd, Alijan Tabarraei. Expression and purification of HCMV glycoprotein B full protein in Escherichia coli. J Complement Med Res. 2020; 11(3): 60-67. doi:10.5455/jcmr.2020.11.03.08

Web Style

Somayeh Azadfar, Ahad Yamchi, Ahmad Majd, Alijan Tabarraei. Expression and purification of HCMV glycoprotein B full protein in Escherichia coli. http://www.jocmr.com/?mno=10846 [Access: October 11, 2020]. doi:10.5455/jcmr.2020.11.03.08

AMA (American Medical Association) Style

Somayeh Azadfar, Ahad Yamchi, Ahmad Majd, Alijan Tabarraei. Expression and purification of HCMV glycoprotein B full protein in Escherichia coli. J Complement Med Res. 2020; 11(3): 60-67. doi:10.5455/jcmr.2020.11.03.08

Vancouver/ICMJE Style

Somayeh Azadfar, Ahad Yamchi, Ahmad Majd, Alijan Tabarraei. Expression and purification of HCMV glycoprotein B full protein in Escherichia coli. J Complement Med Res. (2020), [cited October 11, 2020]; 11(3): 60-67. doi:10.5455/jcmr.2020.11.03.08

Harvard Style

Somayeh Azadfar, Ahad Yamchi, Ahmad Majd, Alijan Tabarraei (2020) Expression and purification of HCMV glycoprotein B full protein in Escherichia coli. J Complement Med Res, 11 (3), 60-67. doi:10.5455/jcmr.2020.11.03.08

Turabian Style

Somayeh Azadfar, Ahad Yamchi, Ahmad Majd, Alijan Tabarraei. 2020. Expression and purification of HCMV glycoprotein B full protein in Escherichia coli. Journal of Complementary Medicine Research, 11 (3), 60-67. doi:10.5455/jcmr.2020.11.03.08

Chicago Style

Somayeh Azadfar, Ahad Yamchi, Ahmad Majd, Alijan Tabarraei. "Expression and purification of HCMV glycoprotein B full protein in Escherichia coli." Journal of Complementary Medicine Research 11 (2020), 60-67. doi:10.5455/jcmr.2020.11.03.08

MLA (The Modern Language Association) Style

APA (American Psychological Association) Style

Somayeh Azadfar, Ahad Yamchi, Ahmad Majd, Alijan Tabarraei (2020) Expression and purification of HCMV glycoprotein B full protein in Escherichia coli. Journal of Complementary Medicine Research, 11 (3), 60-67. doi:10.5455/jcmr.2020.11.03.08

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