Publication:
High level of soluble expression in Escherichia coli and characterisation of the CyaA pore-forming fragment from a Bordetella pertussis Thai clinical isolate

Issued Date

2008-02-01

Resource Type

Article

ISSN

03028933

DOI

10.1007/s00203-007-0302-1

Other identifier(s)

2-s2.0-38049155783

Rights

Mahidol University

Rights Holder(s)

SCOPUS

Bibliographic Citation

Archives of Microbiology. Vol.189, No.2 (2008), 169-174

Suggested Citation

Busaba Powthongchin, Chanan Angsuthanasombat High level of soluble expression in Escherichia coli and characterisation of the CyaA pore-forming fragment from a Bordetella pertussis Thai clinical isolate. Archives of Microbiology. Vol.189, No.2 (2008), 169-174. doi:10.1007/s00203-007-0302-1 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/19373

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Title

High level of soluble expression in Escherichia coli and characterisation of the CyaA pore-forming fragment from a Bordetella pertussis Thai clinical isolate

Author(s)

Busaba Powthongchin
 Chanan Angsuthanasombat

Other Contributor(s)

Mahidol University
 Silpakorn University

Abstract

Bordetella pertussis adenylate cyclase toxin-haemolysin (CyaA) can permeabilise erythrocytes by forming lytic pores. Here, a gene segment encoding CyaA pore-forming (CyaA-PF) domain cloned from genomic DNA of B. pertussis Thai isolate was over-expressed in Escherichia coli as a 126-kDa soluble protein which cross-reacted with anti-RTX monoclonal antibody. By co-expressing with acyltransferase CyaC, the CyaA-PF protein was found palmitoylated at Lys983. Unlike E. coli lysate with the non-acylated form, the lysate containing acylated CyaA-PF exhibited high haemolytic activity against sheep erythrocytes. This study presents that the recombinant CyaA-PF protein comprising pore-forming domain can be expressed separately as soluble native-folded precursor that conserves at least part of its functionality. © 2007 Springer-Verlag.

Keyword(s)

Immunology and Microbiology

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URI

https://repository.li.mahidol.ac.th/handle/123456789/19373

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