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Purification of recombinant budgerigar fledgling disease virus VP1 capsid protein and its ability for in vitro capsid assemblyA recombinant system for the major capsid VP1 protein of budgerigar fledgling disease virus has been established. The VP1 gene was inserted into a truncated form of the pFlag-1 vector and expressed in Escherichia coli. The budgerigar fledgling disease virus VP1 protein was purified to near homogeneity by immunoaffinity chromatography. Fractions containing highly purified VP1 were pooled and found to constitute 3.3% of the original E. coli-expressed VP1 protein. Electron microscopy revealed that the VP1 protein was isolated as pentameric capsomeres. Electron microscopy also revealed that capsid-like particles were formed in vitro from purified VP1 capsomeres with the addition of Ca2+ ions and the removal of chelating and reducing agents.
Document ID
20050000353
Document Type
Reprint (Version printed in journal)
Authors
Rodgers, R. E. (Kansas State University Manhattan 66506)
Chang, D.
Cai, X.
Consigli, R. A.
Spooner, B. S.
Date Acquired
August 22, 2013
Publication Date
May 1, 1994
Publication Information
Publication: Journal of virology
Volume: 68
Issue: 5
ISSN: 0022-538X
Subject Category
Life Sciences (General)
Funding Number(s)
CONTRACT_GRANT: CA07319
Distribution Limits
Public
Copyright
Other
Keywords
NASA Discipline Developmental Biology
Non-NASA Center