1887

Abstract

Summary: pBF4 is a 41 kb conjugative R-plasmid that confers MLS (macrolide-lincosamide-streptogramin B) resistance in spp. To identify pBF4 genes governing conjugation, recombinational mutagenesis using a suicide vector carrying fragments of the pBF4 plasmid was employed. One of six independent insertion mutants of pBF4 isolated using this method was found to be conjugation-deficient. Nucleotide sequence analysis around the insertion site on this plasmid revealed a 2.8 kb ORF that encoded a putative 110 kDa protein. A corresponding protein was observed when a 12 kb DNA fragment containing this ORF was used to program an transcription-translation system. Both the ORF and the predicted protein were novel when compared to available database sequences. This gene was designated (). Polyclonal rabbit antibodies that recognized a sub-sequence polypeptide of BctA reacted with a 55 kDa protein in Western blot analysis using a total protein extract from containing pBF4. The protein was not present in a strain containing the conjugation-deficient insertion mutant of pBF4. The 55 kDa protein was associated with the membrane fraction of Although the cellular and biochemical basis of -promoted conjugation remains unknown, this work demonstrates the existence of a heretofore unrecognized gene in bacterial conjugation, and the mutagenesis system used provides the means to isolate and characterize other genes involved in conjugal transfer in spp.

Funding
This study was supported by the:
  • US PHS (Award R01 DE04224)
  • P50 (Award DE10703)
  • National Institute for Dental Research
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/content/journal/micro/10.1099/00221287-143-7-2155
1997-07-01
2021-10-27
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