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Expression of an EAF plasm id\u2010encoded type IV fimbria, the bundle\u2010forming pilus (BFP), is associated with the LA phenotype. Tn<jats:italic>phoA<\/jats:italic> insertions in <jats:italic>bfpA<\/jats:italic>, the gene encoding the major structural subunit of the BFP, abolish LA. While <jats:italic>bfpA::<\/jats:italic>Tn<jats:italic>phoA<\/jats:italic> mutants cannot be complemented for LA by plasmids carrying the <jats:italic>bfpA<\/jats:italic> gene alone <jats:italic>in trans<\/jats:italic>, this work shows that they can be complemented by plasmids carrying the <jats:italic>bfpA<\/jats:italic> gene, as well as approximately 10kb of downstream sequence, suggesting that such mutations have polar effects on downstream genes. The identification and characterization of a cluster of 13 genes immediately downstream of <jats:italic>bfpA<\/jats:italic> are described. The introduction into a laboratory <jats:italic>Escherichia coli<\/jats:italic> strain of a plasmid containing these 14 <jats:italic>bfp<\/jats:italic> gene cluster genes, along with pJPN14, a plasmid containing another fragment derived from the EAF plasmid, confers LA ability and BFP biogenesis. However, when a mutation is introduced into the last gene of the <jats:italic>bfp<\/jats:italic> cluster, neither LA nor BFP biogenesis is conferred. This work also provides evidence to show that the fragment cloned in pJPN14 encodes a factor(s) which results in increased levels of the pilin protein. Finally, it is shown that expression of the 14 genes in the <jats:italic>bfp<\/jats:italic> cluster from an IPTG\u2010inducible promoter, in the absence of pJPN14, is sufficient to reconstitute BFP biogenesis in a laboratory <jats:italic>E. cob<\/jats:italic> strain, but is insufficient for LA. This is the first report demonstrating the reconstitution of a type IV pilus in a laboratory <jats:italic>E. coli<\/jats:italic> strain with a defined set of genes. The 8FP system should prove to be a useful model for studying the molecular mechanisms of type IV pilus biogenesis.<\/jats:p>","DOI":"10.1111\/j.1365-2958.1996.tb02620.x","type":"journal-article","created":{"date-parts":[[2006,10,6]],"date-time":"2006-10-06T10:55:59Z","timestamp":1160132159000},"page":"325-337","source":"Crossref","is-referenced-by-count":242,"title":["A cluster of fourteen genes from enteropathogenic <i>Escherichia coli<\/i> is sufficient for the biogenesis of a type IV pilus"],"prefix":"10.1111","volume":"20","author":[{"given":"Kelly D.","family":"Stone","sequence":"first","affiliation":[]},{"given":"Hong\u2010Zhong","family":"Zhang","sequence":"additional","affiliation":[]},{"given":"Lisa K.","family":"Carlson","sequence":"additional","affiliation":[]},{"given":"Michael 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