Selection and characterization of randomly produced mutants of gene V protein of bacteriophage M13

A. P M Stassen, G. J R Zaman, Jan Van Deursen, Schoenmakers, R. N H Konings

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Gene V protein of bacteriophage Ff (M13, f1, fd) is a master regulator of phage DNA replication and phage mRNA translation. It exerts these two functions by binding to single-stranded viral DNA or to specific sequences in the 5' ends of its target mRNAs, respectively. To study the structure/function relationship of gene V protein, M13 gene V was inserted in a phagemid expression vector and a library of missense and nonsense mutants was constructed by random chemical mutagenesis. Phagemids encoding gene V proteins with decreased biological activities were selected at the nucleotide sequences of their gene V fragments were determined. Furthermore, the mutant proteins were characterized both with respect to their ability to inhibit the production of phagemid DNA transducing particles and their ability to repress the translation of a chimeric lacZ receptor gene whose expression is controlled by the promoter and translational initiation signals of M13 gene II. From the data obtained, it can be deduced that the mechanism by which gene V protein binds to single-stranded DNA differs from the mechanism by which it binds to its target sequences in the gene II mRNA.

Original languageEnglish (US)
Pages (from-to)1003-1014
Number of pages12
JournalEuropean Journal of Biochemistry
Volume204
Issue number3
StatePublished - 1992
Externally publishedYes

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Genes
Single-Stranded DNA
Bacteriophages
Messenger RNA
Proteins
Lac Operon
Viral DNA
Protein Biosynthesis
Mutant Proteins
DNA Replication
Mutagenesis
Libraries
Gene encoding
DNA
Bioactivity
Gene Expression
Gene expression
Enterobacteria phage M13 gene 5 protein
Nucleotides

ASJC Scopus subject areas

  • Biochemistry

Cite this

Stassen, A. P. M., Zaman, G. J. R., Van Deursen, J., Schoenmakers, & Konings, R. N. H. (1992). Selection and characterization of randomly produced mutants of gene V protein of bacteriophage M13. European Journal of Biochemistry, 204(3), 1003-1014.

Selection and characterization of randomly produced mutants of gene V protein of bacteriophage M13. / Stassen, A. P M; Zaman, G. J R; Van Deursen, Jan; Schoenmakers; Konings, R. N H.

In: European Journal of Biochemistry, Vol. 204, No. 3, 1992, p. 1003-1014.

Research output: Contribution to journalArticle

Stassen, A. P M ; Zaman, G. J R ; Van Deursen, Jan ; Schoenmakers ; Konings, R. N H. / Selection and characterization of randomly produced mutants of gene V protein of bacteriophage M13. In: European Journal of Biochemistry. 1992 ; Vol. 204, No. 3. pp. 1003-1014.
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