Linked Life Data

16896528

Source:http://linkedlifedata.com/resource/pubmed/id/16896528

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
6
pubmed:dateCreated
2006-11-19
pubmed:abstractText
We report the identification and characterization of the single-stranded DNA-binding protein (SSB) from the mesophile and highly radiation-resistant Deinococcus radiopugnans (DrpSSB). PCR-derived DNA fragment containing the complete structural gene for DrpSSB protein was cloned and expressed in Escherichia coli. The gene consisting of an open reading frame of 900 nucleotides encodes a protein of 300 amino acids with a calculated molecular weight of 32.45 kDa and pI 5.34. The amino acids sequence exhibits 43, 44, 79 and 18% identity with Thermus aquaticus, Thermus thermophilus, Deinococcus radiodurans and E. coli SSBs, respectively. The DrpSSB includes two OB folds per monomer and functions as a homodimer. In fluorescence titrations with poly(dT), DrpSSB bound 24-31 nt depending on the salt concentration, and fluorescence was quenched by about 80%. In a complementation assay in E. coli, DrpSSB took over the in vivo function of EcoSSB. The half-lives of DrpSSB were 120 min at 90 degrees C, 60 min at 95 degrees C and 30 min at 100 degrees C. These results were surprising in the context of half-life of SSB from thermophilic T. aquaticus, which has only 30 s of half-life at 95 degrees C. DrpSSB is the most thermostable SSB-like protein identified to date, offering an attractive alternative for TaqSSB and TthSSB in their applications for molecular biology methods and analytical purposes.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
1431-0651
pubmed:author
pubmed:issnType
Print
pubmed:volume
10
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
607-14
pubmed:meshHeading
pubmed-meshheading:16896528-Amino Acid Sequence, pubmed-meshheading:16896528-Bacterial Proteins, pubmed-meshheading:16896528-Cloning, Molecular, pubmed-meshheading:16896528-DNA, Single-Stranded, pubmed-meshheading:16896528-DNA-Binding Proteins, pubmed-meshheading:16896528-Deinococcus, pubmed-meshheading:16896528-Dimerization, pubmed-meshheading:16896528-Escherichia coli, pubmed-meshheading:16896528-Escherichia coli Proteins, pubmed-meshheading:16896528-Genetic Complementation Test, pubmed-meshheading:16896528-Molecular Sequence Data, pubmed-meshheading:16896528-Molecular Weight, pubmed-meshheading:16896528-Protein Denaturation, pubmed-meshheading:16896528-Protein Folding, pubmed-meshheading:16896528-Sequence Analysis, Protein, pubmed-meshheading:16896528-Sequence Homology, Amino Acid, pubmed-meshheading:16896528-Temperature
pubmed:year
2006
pubmed:articleTitle
A highly thermostable, homodimeric single-stranded DNA-binding protein from Deinococcus radiopugnans.
pubmed:affiliation
Department of Microbiology, Gda?sk University of Technology, ul. Narutowicza 11/12, 80-952 Gda?sk, Poland.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't