ABSTRACT
UV-sensitive mutant strain of Haemophilus influenzae Rd MBH3, is 20 times more sensitive to UV irradiation than the wild type strain. The mutation responsible for increased UV sensitivity of the strain was identified as G --> A transition predicting synthesis of truncated UvrAdeltaC44 protein (Balsara & Joshi). Recombinant UvrAdeltaC44 protein was purified for the first time under denaturing conditions. The molecular weight of the recombinant protein was estimated as approximately100 kDa. Recombinant UvrAdeltaC44 protein was found to be less efficient in its ATPase and DNA binding activity as compared to the wild type protein. Recombinant plasmid carrying uvrAdeltaC44 gene could partially complement the UvrA deficiency in E. coli UvrA mutant.
Subject(s)
Adenosine Triphosphatases/chemistry , Bacterial Proteins/chemistry , Base Sequence , Cloning, Molecular , DNA Repair , DNA, Bacterial/genetics , DNA-Binding Proteins/chemistry , Escherichia coli/genetics , Escherichia coli Proteins/genetics , Genes, Bacterial , Genetic Complementation Test , Haemophilus influenzae/genetics , Molecular Weight , Radiation Tolerance , Recombinant Proteins/chemistry , Sequence Deletion , Ultraviolet RaysABSTRACT
Using DNA clones, the physical distance between the linked genes nov and str in Haemophilus influenzae was estimated. Although none of the cloned inserts contained both the markers, pJ1-8StrR 13 (insert of 18·7 kb) included str gene at one end and part of nov gene at the other end of the insert. By EcoRI restriction analysis and by Southern hybridization, the distance between the two EcoRI sites, cutting at which inactivates the two genes, was estimated to be 17·7 kb. A single continuous EcoRI fragment (containing 4 EcoRI sites within it) carrying both the genes intact would need to be 20·4 kb in size. These estimates were confirmed independently using different clones of novr and strr alleles as probes for hybridization with BamHI-digested chromosomal DNA.