High and compact formation of baculoviral polyhedrin-induced inclusion body by co-expression of baculoviral FP25 in Escherichia coli.
Previously, we found that baculoviral polyhedrin (Polh) can successfully be used in Escherichia coli as a fusion partner for the expression of special foreign proteins as inclusion bodies, and the resulting, easily isolatable Polh-induced fusion inclusion bodies had almost the same characteristics as the native Polh. Here, we investigated the effects of co-expression of baculoviral FP25 protein on Polh-induced inclusion-body production in an E. coli expression system, as FP25 is known to be involved specifically in polyhedra formation. Using several analytical tools, including SDS-PAGE, pronase proteolysis, solubilization under alkaline conditions, and electron microscopy, we found that co-expressed FP25 was associated with Polh-induced inclusion bodies and that its co-expression led to formation of compact inclusion bodies as well as high production levels. We confirmed that FP25 co-expression induced higher production levels of other heterologous protein, antimicrobial peptide Hal18, fused with aggregation-prone Polh. Therefore, co-expression of baculoviral FP25 can be promisingly used to increase the levels of baculoviral Polh-fused foreign proteins, especially harmful proteins, expressed as inclusion bodies in an E. coli expression system.
Li
Lin
L
Division of Molecular and Life Sciences, Pohang University of Science and Technology, Pohang 790-784, Korea.
Kim
Young Soo
YS
Hwang
Dong Soo
DS
Seo
Jeong Hyun
JH
Jung
Hee Jung
HJ
Du
Juan
J
Cha
Hyung Joon
HJ
D016367:Baculoviridae; D004926:Escherichia coli / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D007181:Inclusion Bodies, Viral / Q000378:metabolism*; D019590:Nucleocapsid Proteins / Q000008:administration & dosage*; D015202:Protein Engineering / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D014759:Viral Envelope Proteins / Q000378:metabolism*
D019590:Nucleocapsid Proteins; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins