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Production and characterization of recombinant dengue virus type 4 envelope domain III protein

Paper ID Volume ID Publish Year Pages File Format Full-Text
24779 43536 2008 9 PDF Available
Title
Production and characterization of recombinant dengue virus type 4 envelope domain III protein
Abstract

Dengue fever, a mosquito-borne viral disease has become a major worldwide public health problem with a dramatic expansion in recent years. Cultivation process for production of recombinant dengue virus type 4 envelope domain III (rDen 4 EDIII) protein in Escherichia coli was developed for its diagnostic use as well as for further studies in immunoprophylaxis. The dissolved oxygen level was maintained at 20–30% of air saturation. The culture was induced with 1 mM of isopropyl β-d-thiogalactoside when dry cell weight was 13.78 g l−1 and cells were further grown for 4 h to reach 17.31 g l−1 of culture. The protein was overexpressed in the form of insoluble inclusion bodies. The rDen 4 EDIII protein was purified by affinity chromatography and analyzed by SDS-PAGE. The final yield of purified rDen 4 EDIII protein in this method was approximately 196 mg l−1 of culture. The purified protein was recognized in Western blot analysis and enzyme-linked immunosorbent assay (ELISA) with dengue infected human serum samples. These results show that the product has the potential to be used for the diagnosis of dengue infection or for further studies in vaccine development. This production system may also be suitable for the high yield of other recombinant dengue proteins.

Keywords
Cultivation; Dengue; Diagnostic test; ELISA; Escherichia coli
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Production and characterization of recombinant dengue virus type 4 envelope domain III protein
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 134, Issues 3–4, 30 April 2008, Pages 278–286
Authors
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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