PMID: 2499656Jun 1, 1989Paper

The construction and characterization of Neisseria gonorrhoeae lacking protein III in its outer membrane

The Journal of Experimental Medicine
L M WetzlerJ M Koomey

Abstract

Protein III (PIII) is a highly conserved, antigenically stable gonococcal outer membrane protein that is closely associated with the major outer membrane protein, protein I (PI). We have previously reported the cloning of the PIII gene. This gene was inserted into the Eco RI site of the runaway plasmid pMOB45. The beta-lactamase (beta la) Bam HI restriction fragment from the gonococcal plasmid pFA3 was inserted at the Xba I site in the PIII gene. The plasmid construct was Hae III methylated and the PIII/beta la insert was excised with Eco RI and used to transform gonococcal strain F62. One beta la+, ampicillin-resistant transformant was isolated and designated 2D. A Western blot of 2D whole cell lysate was probed with affinity-purified polyclonal PIII antisera. No PIII reactivity was detected. Southern blot analysis was performed on F62 and 2D chromosomal DNA that were cut with Eco RI or Cla I. A PIII DNA probe hybridized with fragments 2.2 kb larger in strain 2D than strain F62. This corresponds to the size of the beta la insert. A beta la-specific probe hybridized with the same 2D restriction fragments as above, but did not react with any F62 fragments, confirming that homologous recombination had occurred. There were minimal...Continue Reading

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Citations

Dec 1, 1995·Trends in Microbiology·M S Blake, L M Wetzler
Nov 7, 2007·Infection and Immunity·Lisa A LewisPeter A Rice
Jun 1, 1992·The Journal of Clinical Investigation·M S Cohen, P F Sparling
Sep 10, 2017·Annual Review of Microbiology·Peter A RiceAnn E Jerse

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