High and Compact Formation of Baculoviral. Polyhedrin-Induced Inclusion Body by Co-Expression of Baculoviral. FP25 in Escherichia coli
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- Title
- High and Compact Formation of Baculoviral. Polyhedrin-Induced Inclusion Body by Co-Expression of Baculoviral. FP25 in Escherichia coli
- Authors
- Li, L; Kim young soo; Hwang, DS; Seo Junh hyun; Jung Hee Jung; Wuan Du; Cha Hyung Joon
- Date Issued
- 2007-04-15
- Publisher
- Wiley
- 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 FP-25 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.
- Keywords
- baculoviaral PP25; baculovinal polyhedrin; inclusion body; fusion protein; Escherichia coli; GREEN-FLUORESCENT PROTEIN; AUTOGRAPHA-CALIFORNICA NUCLEOPOLYHEDROVIRUS; OCCLUSION-DERIVED VIRUS; NUCLEAR-LOCALIZATION; FUSION PARTNER; FT-IR; BODIES; EXPRESSION; GENE; MUTATIONS
- URI
- https://oasis.postech.ac.kr/handle/2014.oak/25695
- DOI
- 10.1002/BIT.21193
- ISSN
- 0006-3592
- Article Type
- Article
- Citation
- BIOTECHNOLOGY AND BIOENGINEERING, vol. 96, no. 6, page. 1183 - 1190, 2007-04-15
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