Year-end Sale % OFF 
Abstract
Enterotoxigenic Escherichia coli (ETEC) is the most significant agent leading to childhood diarrhea and death in developing countries. This bacterium is the cause of 380 thousand deaths in children under five years of age. Due to its prevalence as well as difficulties in its treatment, designing effective vaccines against ETEC is a goal of the World Health Organization (WHO). The Colonization factor B (CfaB) as major subunit of fimberiae has a critical role in bacterial attachment to small intestine epithelium. Hence, the molecule alone or together with other candidate molecules has been considered in vaccine design. In this work, we produced recombinant CfaB in E. coli with the aim of studying its immunogenicity as a component of vaccine. The cfaB gene was cloned into pET28a and regarding the presence of rare codons in cfaB gene, it was not expressed. Therefore, an optimized gene with codon preferences was synthesized and cloned into pET28a vector and subsequently expressed. The recombinant protein was purified with Ni-NTA column and used as an antigen for mice immunization and in ELISA test. Microplate agglutination inhibition test was utilized to show antibody blocking activity. In conclusion, codon optimization is a useful approach for obtaining large quantities of a desired protein. Relying on agglutination inhibition experiment, anti-CfaB serum was able to block the binding of colonization factor antigen I (CFA/I) fimbriated ETEC to erythrocytes. Key words: Enterotoxigenic Escherichia coli (ETEC), colonization factor antigen I, expression, agglutination inhibition
Highlights
Enteric pathogens constitute a serious pediatric threat in less developed countries (Walker, 2005)
Relying on agglutination inhibition experiment, anti-Colonization factor B (CfaB) serum was able to block the binding of colonization factor antigen I (CFA/I) fimbriated Enterotoxigenic Escherichia coli (ETEC) to erythrocytes
The native form of CfaB gene without promoter and signal peptide sequences encodes a mature protein of 147 amino acid residues with a molecular weight of 16.2 kDa
Summary
Enteric pathogens constitute a serious pediatric threat in less developed countries (Walker, 2005). Enterotoxigenic Escherichia coli (ETEC) is among the most frequent bacterial causes of diarrhea (Qadri et al, 2005) and infects adults from industrialized countries visiting these regions, it is known as travelers’ disease (Stauffer et al, 2002). This organism accounts for over 200 million episodes of illness and annually. Adherence and colonization in host cell via filamentous surface structures known as colonization factors (CFs) is a crucial step in ETEC pathogenesis (Anantha et al, 2004). More than 25 human ETEC colonization factors have been identified so far (Fleckenstein et al, 2010); up to 70% of strains
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
