Abstract：
:Escherichia coli secreted protein A (EspA) is a component of the type 3 secretion system (T3SS). The high level of expression when self-stimulated suggests that EspA may be used as a fusion partner. In the present study, EspA was used as a "fusion partner" to construct a fusion expression system, pEspA, in order to improve the expression and solubility of proteins from prokaryotes and eukaryotes. Target proteins were linked to the C-terminus of EspA by a linker containing a YAPQDP sequence, multiple cloning sites and an enterkinase cleavage site. Six proteins, IL-24, Stx2A1, Stx2B, S1, IntiminC300 and GFP, were expressed as EspA-fusion proteins using this vector. The expression level of each protein was enhanced by EspA and the majority of them (Stx2B, IntiminC300, GFP, Stx2A1, IL-24) were expressed in soluble form. EspA-fusion proteins can be purified by affinity chromatography (Sepharose chelated with EspA-specific monoclonal antibody) and by Ni(2+) affinity chromatography for they contain a 6× His tag at their C-terminus. In addition, IL-24 remains soluble and demonstrates certain anti-tumor activity after the removal of EspA by enterkinase. The EspA fusion expression system was efficient in enhancing expression levels and the solubility of target proteins.
journal_name
J Biotechnoljournal_title
Journal of biotechnologyauthors
Cheng Y,Gu J,Wang HG,Yu S,Liu YQ,Ning YL,Zou QM,Yu XJ,Mao XHdoi
10.1016/j.jbiotec.2010.09.940subject
Has Abstractpub_date
2010-11-01 00:00:00pages
380-8issue
3eissn
0168-1656issn
1873-4863pii
S0168-1656(10)01849-3journal_volume
150pub_type
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