Anda di halaman 1dari 18

_________________________________________________________

_________________________________________________________________________

Proteus mirabilis
gram negative, facultative anaerobe rod
causes infections in human urinary tract
(pyelonephritis, kidney stones) followed by
septicaemia
motile bacterium with peritrichous flagella
Morphology change in swarming process

_________________________________________________________________________

Swarming proteus mirabilis


bulls-eye swarming on solid surfaces & self
identity

_________________________________________________________________________

Boundary between P. mirabilis strain


BB2000 and HI4320

_________________________________________________________________________

Aim of study
Identification and characterization of
the genetic mechanism that is
responsible for bacterial self identity

_________________________________________________________________________

Construction of p. mirabilis
mutant library
randomized Tn5-transposon mutagenesis of
rifampicin resistent p. miarabilis BB2000
Tn5-cat

Shuttle vector

cat

conjugal transfer by mating of E.coli [pUTminiTn5Cm] and BB2000

_________________________________________________________________________
tnp: Transposase; mobRP4: conjugal transfer domain

Loss of self recognition I


Screening of 3600 transposon BB2000 mutants
revealed one mutant with phenotypic loss of self
recognition
identification of self (Ids)

_________________________________________________________________________

Loss of self recognition II


six gene cluster idsABCDEF carries information for
identification of self

_________________________________________________________________________

Phenotyping of individual
idsABCDEF knock outs

idsB idsF are required for self recognition !


_________________________________________________________________________

Phenotyping of individual
idsABCDEF knock outs in
HI4320 background

idsB,
idsD and
idsCidsE
and idsF
are essential
can be complemented
for self recognition
by allels
! of different
_________________________________________________________________________

idsD and idsE encode identity


determinants

.but the ids gene cluster is not the sole determinant of boundary
ation !

boundary between BB2000+pidsHI4320 and HI4320+pidsBB2000


_________________________________________________________________________

Conserved organization of ids


gene cluster in other P.
mirabilis isolates

idsABCDEF with nearly identical length and ~96% identity to homolog

idsD and idsE have variable domains in protein sequences


_________________________________________________________________________

Additional putative idsE genes


in genome of P. mirabilis
HI4320

_________________________________________________________________________

Model of self recognitation

idsA is not required for self identity; idsF overtakes function distinct from

dsB
_________________________________________________________________________
and idsC in self-nonself recognition; idsD and idsE are molecular self
dentifiers

Summary
gene locus idsABCDEF of P. mirabilis is important for
self-nonself discrimination and boundary formation
Conserved ids gene cluster has been found in other
proteus mirabilis strains
idsA is not required for parental recognition
idsB, idsC and idsF are interchangeable between
strains
idsD and idsE are specific molecular identifiers that
possess strain dependent variable domains
other ids homologs in bacterial genome may
contribute to the self recognition process

_________________________________________________________________________

Thank you for your attention

_________________________________________________________________________

P. mirabilis boundaries

_________________________________________________________________________

Transposition and pBBR-MCS2


shuttle vector for mutagenesis

_________________________________________________________________________

Anda mungkin juga menyukai