Cloning of the Genes for AF/R1 Pili from Rabbit Enteroadherent Escherichia coli RDEC-1 and DNA Sequence of the Major Structural Subunit
Abstract
AF/R1 pili on the surface of Escherichia coli RDEC-1 promote attachment of the bacteria to rabbit intestinal brush borders. In order to characterize AF/R1 pili and manipulate their expression, we cloned the genes necessary for AF/R1 expression; determined the size of proteins produced in minicells; located the gene encoding the major structural subunit, named AfrA; and determined the DNA sequence of afrA as well as the sequence of 700 additional nucleotides upstream of afrA. Two contiguous EcoRI fragments spanning 7.9 kilobases were cloned from the 86-megadalton plasmid of RDEC-1 into vector pUC19 to make plasmid pW1. Bacteria carrying pW1 produced AF/R1 pili that were recognized by AF/R1-specific antiserum and promoted adherence of bacteria to brush borders prepared from rabbit intestine. Proteins with a molecular weight of 17,000 (17K proteins), which was the size of AfrA, as well as 15K, 15.5K, 26K, 28K, and 80K proteins were detected in minicells carrying pW1.
Document Details
- Document Type
- Technical Report
- Publication Date
- Apr 01, 1990
- Accession Number
- ADA224135
Entities
People
- Edgar C. Boedeker
- Marcia K. Wolf
Organizations
- Walter Reed Army Institute of Research