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Journal of Bacteriology, November 2001, p. 6274-6281, Vol. 183, No. 21
Department of Molecular and Cell Biology,
Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutsumi-dori Amamiya-machi, Aoba-ku, Sendai
981-8555,1 and Department of
Microbiology, Miyazaki Medical College, 5200 Kiyotake, Miyazaki
899-1692,2 Japan
Received 16 April 2001/Accepted 31 May 2001
Carotovoricin Er is a phage-tail-like bacteriocin produced by
Erwinia carotovora subsp. carotovora
strain Er, a causative agent for soft rot disease in plants. Here we
studied binding and killing spectra of carotovoricin Er preparations
for various strains of the bacterium (strains 645Ar, EC-2, N786, and
P7) and found that the preparations contain two types of carotovoricin Er with different host specificities; carotovoricin Era possessing a
tail fiber protein of 68 kDa killed strains 645Ar and EC-2, while
carotovoricin Erb with a tail fiber protein of 76 kDa killed strains
N786 and P7. The tail fiber proteins of 68 and 76 kDa had identical
N-terminal amino acid sequences for at least 11 residues. A search of
the carotovoricin Er region in the chromosome of strain Er indicated
the occurrence of a DNA inversion system for the tail fiber protein
consisting of (i) two 26-bp inverted repeats inside and downstream of
the tail fiber gene that flank a 790-bp fragment and (ii) a putative
DNA invertase gene with a 90-bp recombinational enhancer sequence. In
fact, when a 1,400-bp region containing the 790-bp fragment was
amplified by a PCR using the chromosomal DNA of strain Er as the
template, both the forward and the reverse nucleotide sequences of the
790-bp fragment were detected. DNA inversion of the 790-bp fragment
also occurred in Escherichia coli DH5
0021-9193/01/$04.00+0 DOI: 10.1128/JB.183.21.6274-6281.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
DNA Inversion in the Tail Fiber Gene Alters the Host Range
Specificity of Carotovoricin Er, a Phage-Tail-Like Bacteriocin of
Phytopathogenic Erwinia carotovora subsp.
carotovora Er
when two
compatible plasmids carrying either the 790-bp fragment or the
invertase gene were cotransformed into the bacterium. Furthermore,
hybrid carotovoricin CGE possessing the tail fiber protein of 68 or 76 kDa exhibited a host range specificity corresponding to that of
carotovoricin Era or Erb, respectively. Thus, a DNA inversion altered
the C-terminal part of the tail fiber protein of carotovoricin Er,
altering the host range specificity of the bacteriocin.
*
Corresponding author. Mailing address: Department of
Molecular and Cell Biology, Graduate School of Agricultural Science, Tohoku University, Aoba-ku, Sendai 981-8555, Japan. Phone:
81(22)717-8779. Fax: 81(22)717-8780. E-mail:
ykamio{at}biochem.tohoku.ac.jp.
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