@article{mbs:/content/journal/micro/10.1099/00221287-143-7-2155, author = "Morgan, Roderick M. and Macrina, Francis L.", title = "bctA: a novel pBF4 gene necessary for conjugal transfer in Bacteroides spp.", journal= "Microbiology", year = "1997", volume = "143", number = "7", pages = "2155-2165", doi = "https://doi.org/10.1099/00221287-143-7-2155", url = "https://www.microbiologyresearch.org/content/journal/micro/10.1099/00221287-143-7-2155", publisher = "Microbiology Society", issn = "1465-2080", type = "Journal Article", keywords = "conjugation", keywords = "recombinationai mutagenesis", keywords = "pBF4", keywords = "bctA", keywords = "Bacteroides fragilis", abstract = "Summary: pBF4 is a 41 kb conjugative R-plasmid that confers MLS (macrolide-lincosamide-streptogramin B) resistance in Bacteroides 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 in vitro transcription-translation system. Both the ORF and the predicted protein were novel when compared to available database sequences. This gene was designated bctA (Bacteroides conjugal transfer). 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 Bacteroides fragilis containing pBF4. The protein was not present in a B. fragilis strain containing the conjugation-deficient insertion mutant of pBF4. The 55 kDa protein was associated with the membrane fraction of B. fragilis. Although the cellular and biochemical basis of bctA-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 Bacteroides spp.", }