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Viability and preliminary in vivo characterization of site directed mutants of Escherichia coli single‐stranded DNA‐binding protein
Author(s) -
Carlini Leslie E.,
Porter Ronald D.,
Curth Ute,
Urbanke Claus
Publication year - 1993
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1993.tb00977.x
Subject(s) - biology , escherichia coli , mutant , dna , microbiology and biotechnology , genetics , computational biology , biochemistry , gene
Summary Site‐directed mutations involving selected amino acids of Escherichia coli single‐stranded DNA‐binding protein (SSB) were tested for their in vivo functionality when introduced into a chromosomal ssb deletion strain on a plasmid. All mutants complemented the ssb deletion for viability when present on a pSC101 derivative. The generation time with ssbW54S doubled in comparison to the ssb * control, and both the ssbW54S‐ and ssbH55K‐containing strains exhibited temperature sensitivity. ssbH55K, ssbW54S, ssbW88T, and ssbH55Y (ssb‐1) strains displayed reduced survival to ultraviolet irradiation, while ssbW40T and ssbF60L strains were comparable to the ssb + control strain. This study represents the first investigation of the in vivo properties of ssb mutations constructed for in vitro analysis of DNA binding by SSB.