Prev. Article Next Article Table of Contents

Article

Expression Profiling of Herpesvirus and Vaccinia Virus Proteins Using a High-Throughput Baculovirus Screening System

Mian Gao,^†^# Nicole Brufatto,^† Tricia Chen,^† Laura Lea Murley,^† Rosanne Thalakada,^† Megan Domagala,^† Bryan Beattie,^† Daniel Mamelak,^† Vicki Athanasopoulos,^‡ David Johnson,^§ Grant McFadden, Christian Burks,^† and Lori Frappier*^†^‡

Affinium Pharmaceuticals, 100 University Avenue, Toronto, Canada M5J 1V6, Department of Medical Genetics and Microbiology, University of Toronto, 1 Kings College Circle, Toronto, Canada M5S 1A8, Department of Molecular Microbiology and Immunology, Oregon Health Sciences University, Portland, Oregon 97201-3098, Department of Microbiology and Immunology, University of Western Ontario, and Robarts Research Institute, 1400 Western Road, London, Ontario, Canada N6G 2V4

J. Proteome Res., 2005, 4 (6), pp 2225–2235

DOI: 10.1021/pr050137u

Publication Date (Web): September 17, 2005

†

Affinium Pharmaceuticals.

Present address: Gene Expression & Protein Biochemistry, Bristol-Myers Squibb Company, P.O. Box 4000, Princeton, NJ 08543.

Present address: CanReg Inc., 4 Innovation Drive, Dundas, Ontario Canada L9H 7P3.^& Present address: Custom Biologics, 1 Marmac Drive, Toronto, Ontario Canada M9W 1E7.

‡

University of Toronto.

Present address: Research School of Biological Sciences, Sullivan's Creek Road, The Australian National University, ACT Acton 2601 Australia.

Oregon Health Sciences University.

University of Western Ontario.

Present address: Ontario Genomics Institute, 149 College Street, suite 500, Toronto, Ontario Canada M5T1P5.

To whom correspondence should be addressed: lori.frappier@utoronto.ca.

Abstract

We have developed a high-throughput system for generating baculoviruses and testing the expression, solubility, and affinity column purification of encoded proteins. We have used this system to generate baculoviruses for and analyze the expression of 337 proteins from three different herpesviruses (HSV-1, EBV, and CMV) and vaccinia virus. Subsets of these proteins were also tested for expression and solubility in E. coli. Comparisons of the results in the two systems are presented for each virus.

Keywords: baculovirus • high-throughput protein expression • Epstein−Barr virus • herpes simplex virus • cytomegalovirus • vaccinia

View: Full Text HTML | Hi-Res PDF