Open AccessThe use of Tn phoA in Erwinia amylovora to generate random fusions of alkaline phosphatase to extracytoplasmic proteins
Author(s) -
Coleman Mark J.,
Milner Jonathan S.,
Cooper Richard M.,
Roberts Ian S.
Publication year - 1991
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1991.tb04655.x
Subject(s) - cell envelope , erwinia , biology , virulence , microbiology and biotechnology , signal peptide , alkaline phosphatase , biochemistry , bacteria , escherichia coli , genetics , gene , peptide sequence , enzyme
Tn phoA has been introduced with high efficiency into the chromosome of the phytopathogenic bacterium Erwinia amylovora . Transposition occurred with apparent randomness and single insertions were predominant. Fusion proteins were detected in cell envelope fractions from PhoA + mutants and a range of alkaline phosphatase activities was onserved. The results provide the first evidence that sequences encoding signal peptides are present in E. amylovora and that Tn phoA may be a valuable tool for the study of the translocation, regulation and function of E. amylovora extracytoplasmic proteins. In particular, Tn phoA mutagenesis should be applicable to the identification of cell envelope proteins involved in the virulence of this organism.