Cloning and Purification of a Unique Lysozyme Produced by Bacillus Phage φ 29

A DNA fragment of the bacteriophage φ 29 chromosome, encoding the entire sequence of φ 29 gene 15, has been cloned into the Escherichia coli expression vector pPLc245 under the control of the phage λ major leftward promoter, PL. Upon heat induction, a protein with an apparent molecular mass of 26 kD...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1987-02, Vol.84 (4), p.955-958
Main Authors: Saedi, Mohammad S., Garvey, Kevin J., Ito, Junetsu
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Amino acids
Bacillus - enzymology
Bacteriophages
Cloning, Molecular
Electrophoresis
Enzymes
Evolution
Gels
Genes
Muramidase - genetics
Muramidase - isolation & purification
Nucleotide sequences
Plasmids
Proteins
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Summary:A DNA fragment of the bacteriophage φ 29 chromosome, encoding the entire sequence of φ 29 gene 15, has been cloned into the Escherichia coli expression vector pPLc245 under the control of the phage λ major leftward promoter, PL. Upon heat induction, a protein with an apparent molecular mass of 26 kDa was overproduced. The molecular mass of this protein corresponds to the 28 kDa predicted for the product of gene 15 from its nucleotide sequence. The overproduced protein has been purified to near homogeneity and confirmed to be the product of gene 15 by amino acid sequence analysis of its N terminus. The purified product of gene 15 has a lysozyme activity similar to other phage-type lysozymes: products of phage T4 gene e and of phage P22 gene 19. However, to our knowledge φ 29 lysozyme is structurally unique among the phage-type lysozymes.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.84.4.955