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987P fimbrial gene identification and protein characterization by T7 RNA polymerase‐induced transcription and TnphoA mutagenesis
Author(s) -
Schifferli D. M.,
Beachey E. H.,
Taylor R. K.
Publication year - 1991
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1991.tb01826.x
Subject(s) - biology , rna polymerase ii , mutagenesis , gene , transcription (linguistics) , rna polymerase , genetics , t7 rna polymerase , polymerase , rna polymerase iii , microbiology and biotechnology , computational biology , rna , gene expression , promoter , mutation , escherichia coli , linguistics , philosophy , bacteriophage
Summary The 987P fimbrial gene cluster has been previously cloned as a 12 kb fragment from prototype strain 987. Gene products encoded by the whole clone were analysed by utilizing an in vivo system based on the induction of transcription by T7 RNA polymerase. The sensitivity of this technique permitted us to identify new proteins involved in 987P fimbriation. In total, eight proteins were detected, their genes ( fasA to fasH ) were mapped and their orientation of transcription determined. Several of the gene products demonstrated typical properties of exported proteins. Precursor and processed forms could be correlated after inhibiting protein transport with ethanol. The detection of enzymatically active fusion proteins after Tn phoA (Tn 5 IS 50 L :: phoA ) mutagenesis supported and complemented these results. One protein encoded by the 12kb fragment was found not to be related to fimbriation but rather the product of the STla gene, identified as a component of a Tn 1681 ‐like transposon.