PMID:7721718

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Citation

Harlow, KW, Nygaard, P and Hove-Jensen, B (1995) Cloning and characterization of the gsk gene encoding guanosine kinase of Escherichia coli. J. Bacteriol. 177:2236-40

Abstract

The Escherichia coli gsk gene encoding guanosine kinase was cloned from the Kohara gene library by complementation of the E. coli gsk-1 mutant allele. The cloned DNA fragment was sequenced and shown to encode a putative polypeptide of 433 amino acids with a molecular mass of 48,113 Da. Minicell analysis established the subunit M(r) as 43,500. Primer extension analysis indicated the presence of an adequate Pribnow box and suggested that the transcript contained a 110-base leader sequence. Strains harboring the gsk gene on multicopy plasmids overexpressed both guanosine and inosine kinase activities. N-terminal sequence and amino acid composition analyses of the 43,500-M(r) polypeptide band confirmed the correct reading frame assignment and the identity of this band as the gsk gene product. Comparison of the amino acid sequence with the protein database revealed similarity to regions of other mononucleotide-utilizing enzymes.

Links

PubMed PMC176874

Keywords

Amino Acid Sequence; Base Sequence; Cloning, Molecular; DNA, Bacterial/genetics; Escherichia coli/enzymology; Escherichia coli/genetics; Gene Expression; Genes, Bacterial; Genetic Complementation Test; Molecular Sequence Data; Molecular Weight; Phosphotransferases (Alcohol Group Acceptor)/chemistry; Phosphotransferases (Alcohol Group Acceptor)/genetics; Plasmids/genetics; Protein Conformation; Restriction Mapping; Sequence Homology, Amino Acid; Transcription, Genetic

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