Expression and properties of bacteriophage T4 gene product 11

A plasmid vector for expression of bacteriophage T4 gene product 11 (gp11) in E. coli cells has been constructed. Gp11 is a baseplate protein that connects short tail fibers providing irreversible adsorption of the virus on a cell. A method based on chromatography on hydroxyapatite has been develope...

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Bibliographic Details
Published in:Biochemistry (Moscow) 2001-02, Vol.66 (2), p.141-146
Main Authors: Kurochkina, L P, Leiman, P G, Venyaminov, S Y, Mesyanzhinov, V V
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Bacteria
Base Sequence
Chromatography, Gel
Circular Dichroism
Cloning, Molecular
DNA, Recombinant
E coli
Electrophoresis, Polyacrylamide Gel
Enzymes
Escherichia coli
Escherichia coli - genetics
Fibers
Molecular Sequence Data
Protein Conformation
Recombinant Proteins - chemistry
Recombinant Proteins - genetics
Recombinant Proteins - isolation & purification
Recombinant Proteins - metabolism
Spectrophotometry, Ultraviolet
Viral Proteins - chemistry
Viral Proteins - genetics
Viral Proteins - isolation & purification
Viral Proteins - metabolism
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Summary:A plasmid vector for expression of bacteriophage T4 gene product 11 (gp11) in E. coli cells has been constructed. Gp11 is a baseplate protein that connects short tail fibers providing irreversible adsorption of the virus on a cell. A method based on chromatography on hydroxyapatite has been developed for purification of recombinant gp11. The protein is active in an in vitro complementation assay and transforms defective phage particles lacking gp11 into infective ones. Gel filtration data suggest that the biologically active protein is a trimer. According to CD spectroscopy and sequence analysis data, the polypeptide chain of gp11 contains not less than 20% alpha-helical segments, about 30% beta-structure, and belongs to the class of alpha/beta structural proteins.
ISSN:0006-2979
1608-3040
DOI:10.1023/A:1002831212462