High and compact formation of baculoviral polyhedrin-induced inclusion body by co-expression of baculoviral FP25 in Escherichia coli

✍ Scribed by Lin Li; Young Soo Kim; Dong Soo Hwang; Jeong Hyun Seo; Hee Jung Jung; Juan Du; Hyung Joon Cha

• Publisher: John Wiley and Sons
• Year: 2007
• Tongue: English
• Weight: 279 KB
• Volume: 96
• Category: Article
• ISSN: 0006-3592
• DOI: 10.1002/bit.21193

✦ Synopsis

Abstract

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. Biotechnol. Bioeng. 2007;96:1183–1190. © 2006 Wiley Periodicals, Inc.