TY - JOUR
T1 - EspA is a novel fusion partner for expression of foreign proteins in Escherichia coli
AU - Cheng, Yan
AU - Gu, Jiang
AU - Wang, Hai guang
AU - Yu, Shu
AU - Liu, Yan qing
AU - Ning, Ya lei
AU - Zou, Quan ming
AU - Yu, Xue jie
AU - Mao, Xu hu
N1 - Funding Information:
This work was supported by the National S&T Major Project Science and Technology (2009ZX09306-006).
PY - 2010/11
Y1 - 2010/11
N2 - 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 Ni2+ 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.
AB - 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 Ni2+ 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.
KW - Escherichia coli
KW - EspA
KW - Fusion partner
KW - Fusion protein
UR - http://www.scopus.com/inward/record.url?scp=78149464373&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=78149464373&partnerID=8YFLogxK
U2 - 10.1016/j.jbiotec.2010.09.940
DO - 10.1016/j.jbiotec.2010.09.940
M3 - Article
C2 - 20854850
AN - SCOPUS:78149464373
SN - 0168-1656
VL - 150
SP - 380
EP - 388
JO - Journal of Biotechnology
JF - Journal of Biotechnology
IS - 3
ER -