Identification and characterization of hitherto unknown Mycoplasma pneumoniae proteins

Mol Microbiol. 1994 Jul;13(2):337-48. doi: 10.1111/j.1365-2958.1994.tb00427.x.

Abstract

Eleven hitherto unknown Mycoplasma pneumoniae proteins were identified and characterized with respect to their size and subcellular location. This was carried out through the construction of in vitro gene fusions between a modified mouse dehydrofolate reductase (dhfr) gene and selected regions (cosmid clones) of the M. pneumoniae genome and expressing them in Escherichia coli. Positive clones were identified using antibodies against specific fractions of M. pneumoniae. The deduced protein sequences of 11 out of 30 clones did not show significant homologies to known proteins in protein data-bank searches. Monospecific antibodies against these 11 fusion proteins were used to determine the size and cellular location of the corresponding M. pneumoniae proteins by immunoscreening Western blots of SDS-acrylamide gels from M. pneumoniae cell extracts.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Proteins / genetics
  • Bacterial Proteins / isolation & purification*
  • Base Sequence
  • Cloning, Molecular
  • Cosmids
  • Detergents / pharmacology
  • Escherichia coli
  • Gene Library
  • Membrane Proteins / isolation & purification
  • Mice
  • Molecular Sequence Data
  • Mycoplasma pneumoniae / chemistry*
  • Mycoplasma pneumoniae / drug effects
  • Mycoplasma pneumoniae / genetics
  • Mycoplasma pneumoniae / ultrastructure
  • Open Reading Frames
  • Recombinant Fusion Proteins
  • Sequence Homology, Nucleic Acid
  • Subcellular Fractions / chemistry
  • Tetrahydrofolate Dehydrogenase / genetics

Substances

  • Bacterial Proteins
  • Detergents
  • Membrane Proteins
  • Recombinant Fusion Proteins
  • Tetrahydrofolate Dehydrogenase

Associated data

  • GENBANK/Z32646
  • GENBANK/Z32647
  • GENBANK/Z32648
  • GENBANK/Z32649
  • GENBANK/Z32650
  • GENBANK/Z32651
  • GENBANK/Z32652
  • GENBANK/Z32653
  • GENBANK/Z32654
  • GENBANK/Z32655
  • GENBANK/Z32656
  • GENBANK/Z32657
  • GENBANK/Z32658
  • GENBANK/Z32659
  • GENBANK/Z32660
  • GENBANK/Z32661
  • GENBANK/Z32662
  • GENBANK/Z32663
  • GENBANK/Z32664
  • GENBANK/Z32665
  • GENBANK/Z32666
  • GENBANK/Z32671