fulltext.study @t Gmail

Efficient expression and purification of porcine circovirus type 2 virus-like particles in Escherichia coli

Paper ID Volume ID Publish Year Pages File Format Full-Text
22742 43380 2016 8 PDF Available
Title
Efficient expression and purification of porcine circovirus type 2 virus-like particles in Escherichia coli
Abstract

•Full-length of PCV2 capsid protein is efficiently expressed in E. coli and proceeds to form virus-like particles (VLPs).•The VLPs can be simply purified by a single-step gel-filtration chromatography.•VLPs-immunized mice elicit both humoral and cell-mediated immune response to PCV2.

Porcine circovirus type 2 (PCV2) capsid (Cap) protein has been successfully used as a vaccine to control porcine circovirus associated disease (PCVAD). Most PCV2 subunit vaccines are recombinant Cap protein expressed in baculovirus/insect cell expression system, but using this eukaryotic system is laborious and expensive. In our previous study, full-length of PCV2Cap protein expressed in Escherichia coli formed virus-like particles (VLPs). This expression system has the advantages of being relatively simple and inexpensive. In this study, we constructed a recombinant plasmid containing the full-length codon-optimized cap (ORF2) gene to improve high-level expression of recombinant Cap protein (rCap) with no changed amino acids. The highly water-soluble rCap protein was purified by a single-column, high-throughput fractionation procedure based on size exclusion chromatography. Yield was 10 mg per 200 ml bacterial culture. The rCap protein self-assembled into VLPs of diameter 25–30 nm that contained exogenous nucleic acids. The immunogenicity of PCV2 VLPs was analyzed by immunizing mice. VLP-immunized mice mounted specific immune responses to PCV2. Thus, expression of rCap in E. coli was feasible for large-scale production of PCV2 VLPs, which could potentially be used for a VLP-based PCV2 vaccine.

Keywords
PCV2 capsid protein; Virus-like particles; Size exclusion chromatography; Vaccine
First Page Preview
Efficient expression and purification of porcine circovirus type 2 virus-like particles in Escherichia coli
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 220, 20 February 2016, Pages 78–85
Authors
, , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us