You are on page 1of 23

05.L2.

20L2

Fenotip

Variabilitate

Prof Lia Monica Junie

Cl*:llrrlrrr Iisptrrlr;*.ti t<r

l)N,\ llrrrrrrrl;c

l.:rr{rr-}'rcr
lls-A Lcpsit

llrl

lotalitatea informafiei genetice exister ri6 intr-o celultr

modificarea informafiei genetice

orilicrt

,,

ff

{f,\.4 lr!s**lri{
"{ssdl3

Ap.rprsib

irrrr

u'"r -1,}r!lf
,

bacteriile detin
n.he.etgIioraqp
t
',*l
(;--\
|
_
l(

'^
$,: ''b

d*J

{}j
T:,

v"riabilitate
cromozom bacterian

'.\

ptasmide

L\-/q
,**9*,
I

:";i;;

al:tb-J;[ !
:""-' s.h''/-

elen,c,.rq genetice

mo6ilitn

:,q,g|}:,,lll

l!*.*t***tat!:

material genetic

':

Alterarea informatiei genetice


Aditia/deletia debazeazome

i,;,.i :'

05.L2.20L2

muultla

functrionale

|J

fneumococlt(^
.

Streptococcus pneumoniae

Et rii.nccapgrlari.,"',*"*,H,fj"tldlr1j3s;dottu

El colonudtlpRpc
rurediilc de cufturi

El

uurtafre

dlr

pllellln(]('or:l

hrilnl

.,1

;nrf'l*1ruim

cirpsulilti S

i i $ilt$io,: ir mfrris r{si$e$l


d espfu r: r'tririues

ftnlsiimi

tr

Trlurforunree

inactivafi prin ctrlduri" 60o

****-*tr
't

TIT.ANSFOI<^^AI
iON
(..
descoperit de Griffith

ln

1928,
virulentei pneumococilor
Streplococcus pneumoniae

'& ',@

:#*#r#HF

fransformarea pneumococului R nepatogen in pneumococ S viu gi


patogerq cu revenirea propriettrlii de a produce colonii S pe
mediile de culturl 9i a capacitigii de patogenitate
l2/5/20t2

htur tonlo

)unl.

05.L2.2012

s' **&qi

s*6ivr

Experiment Griffrth

;''
'r:s.-'
i
''%.T-*
{,D

-di

inocularea subcutanatl a pnumooocilor la goareci

I i'lattrh
It

J--i ::--@
tillk
ii-i-uat*,
'f"r*1
f*V,
f
f*Y
-t-i$gl"
-;*'f*$.,t;k:#"
-il$dl"
-: . I " ' i ryn : ';.r-r.rr

!1"

,
",.,
r-'f,l-.1:i'."

'?

t 'tt*,

t':t',r,,

"';'

:**P .:::*F,::*P::kF
illi:----"

;ffiil;''

n@psulali,

vii,

vii

sepsurariinsrtiwri R+sinactivrt

epsulali cildur[

cl:','*",r "*
ld<erf

$s'

b*.r@!

rl*#f

*i]@6*#,

Ftransformarea pneumococului R nepatogen ln pneumococ S viu gi


patogen, cu revenirea proprieti{ii de a produce colonii S pe mediile de
cdtura fi a capaciti$ii de patogenitate

Deces

pneumococi nepatogeni, avirulenti inoculali Ia goarcce

-goriceii

i'_,(:t

supraviefti-

n&&

!t"'****\..

'l', {'ii

";,,

;f:l:
ii'

''&.r

ffi.

n^

ii,'' \

iy ;

:^

*r"[E-\;.; ffi

.&=

t'l Criflith

Pneumococi inoculat la goareci, decesr 24-48 ore' stl)ticcmie

tr
tr

M#ffYo'***

"principiu traNformant "

::

fr'*<({r{4i'+(ts
lh1...t srilr&

/i\

f;;{}T-.:t .{l1?..j. , "


E
Din sdngele anirnalelor decedate s-a izolal un pneuntococ viu,
tip S, capsulat, patogen

,;.{i . iri t4\

and

r
rFi

,&-,Z v;m
-ffi

Wr

r:

rr

Maclrod

n'^lgj"

-!?,vr,

&ww
Ill

II

il

Transformare R in S virulent

I -t\-rI

i)*'i-;"
*";'

nu a putut explica fenomenul produs

Griffith experiment

'fM,l-

''

tEa

dttrr
- JG

{dr*ar* &

<d':"'
lJ

I allf.rlt u\r

<un*
,r# #9

?#*r-l
D'^;'4tr"rn't

transfer gene DNA la S, pneumoniae

05.r2.20t2

rrr.ildrr*i.r:r
ll...a*:.i$

ir .r-i

l* {r..}* r

I r:e **^.-.*.*-**-,.

:rE::

.:

**-::
;:

*'!/iqla
-,_
\-_

E
E
E

-^

-:f

-3

:*a-.

'_/

fenomengenetic
transfer de ADN liber
de la o bacterie donatoare (D) la o bacterie receptoare

Inlomatii ir)portante rol ADN

E).

exog6note

.-.|
rsaomb ina i$on g6ri6tique

E
E

Fragmentul de AI)N transferat

natural prin autoliza bacteriilor

gi

Schirnb material scnetic

CI fcnonenul

de transforrnareevidenliat

E St r( pt oco ccu s I n eumoniae,


C) r rlre hrctcrii aram pozitive:

fenomenul de trlnsformare

*'-tKuTt-,.il

Cl
E

nStalilococi,
Dgcn Lhcrllus;
lbacil cirbrrnos
l)Nctcrii Grrnr ncgative:
!1le nrofili,

INeisseria
lirrr:i.
- este un fenomen iar lntAlnit in natura- sunt necesare arumite conditii

05.L2.20L2

conditii necesare

E
E
tr
E
E
E

FragmentuldeADN
transferat
dimensiuneaminimllff
Dal
1% dinADN-ul donalor
ConcentratiaADN-uluiin

mediu miriimum 105 ng/ml


o perioadd de contact lnte
ADN $i bacteria receptoare
o densitatea celulelor
reccptoarc in mediu

I Elbe

ar er i'agmcnftrhi dc
.{DN dilr brcterinD.

2..{tirlaleir fi qgnentuhri

.{DN

cle

de bo(terio

',

recept0rre.

m\

[togenote

transformare bactcriana
3. Penetrarea fragmentului de

ADN

ln bacteria R
4. perioada de eclipstr,
5. integrarea ftagrnentului dc ADN
exogen in cromozomul bacteriei
R, prin recombinare omoloaga6. Recombinantul rezultat

li;r:ii'ii:;ai,,."."., .

n\.-.-.

li ''

stadiile transformarii
l)\,\ lJIItrs {{} c}:t,l,

l}:41"1,

DttA

l:ivrfHi

||ocir'('ul,t.

l'!i

II()S',r

I)r.r

05.12.2012

recombinrre omoloaga

Se

produce

o molecula

de

.**/

diferite

-ffie
I molecule dilerite de
I
I

ADN

molecule

sau

circulare

o molecultr circularl
qi fragment de

t^ 'T T^ '1t^
| 1c,,,'rl I ,-,. |

'T

ryt*"s*,*j:i
k@F' \ .r*

ffi?l: ,,.
:'}T*
'"'ao*o*lt

,e,L:
r1r ""

ll*ll*ll
,ll,

fi *:"
*Y.Y*
-^.

,1J.* ,11,

=.**L..*x*rL--**;

ADN
ptrtruns in celule

:.

r....

';u:.-:)r{qe*)::i::*}

.t '

Rr{stHrad Pafentd

}ftA

*fiA

REcnfi?,nerd

ofin

. Proteina ,,rec.A" determinf, gAsirea zonei de omologie intre


fragmentul de ADN exogen (exogenot),
. alinierea exogenotului la endogenot,
. ruperea qi reunirea catenelor gi
. mlegrarea exogenomul m enoogeno! care pnme;Ie gene oe la

recombinare Omoloaga

recombinarea
lntre molecule cu
secvenle omoloage:

I.lEffiil-l.-,..,,/*-*$\...-,,.....
identice/aproape * - .;*

*.,,.1q,,,,

loentlce.

,/
\
I c6terr nucleotide, un*wh*m} '
-.*utr.
-'
|hkc{ofld
I pereclri
\
,..*^J
t'Irtu
Comfrl(mcntafe
r;ts.@
-,s"
|

r pezoncertinse

ffii

'#v;*iir+'

l$i**'@

{trNer'rynd!
it${ixr
Mrn:a

Reparare recombinare
O proteina Rrc

'\

El duplex
O efechrat de

E ADN polimeraza

LaunelebacteriiGram
negative,

I hemofilii,
E
cromozomul bacterian gi ln
- inft"L;;;h,la;-AD].in-.i;i,
Recunoastereasequente
I existtr secvente de ll-14 specifice
baze azotate, care se repetd. H, influenzae:
r 600ori
AAGTGCGGTCA
I la nivelul cirora se produce N. gonothoeae:

1,

:a..

_nnt'\qrri Ijir.nrmrr

\,/

O ligaza

intre celule ale acelessi


specii sau
specii str6ns lnrudite

I i'"',
l{'l

,/o\

l,Fllllwjoiir ''''-

al

t-n"fT{)Tn-n

cauta secYcLtl I r,Iroloaga de

tr

Datoritd recombindni

\o/

legare cattne taiate

rr"r,ftr'rt
,{-^rz
I lr..r..t.L.r..t l r..l.r.

UNA
luslaotz

t''

,",""b\.t\

rfidI]rn
runl.

Pref Xonlo

lrlt-]-:n-rm
!!

05.L2.20L2

N'Iecanismele

transformarii naturale

""bnt"'o- -r.""''"''

fffirliffiiii?
r--*:iYt*'tttt'i)
(}*l::*'-r::l-J
f

{ffi1

Transformarea naturala la bacteriile Gram pozitive

Starea de competen{tr

E'Fffifft{fi:.r
E prezentl du;rr
staFonara

E
E
El

permancnt
(ex. la hcrrrr,lili)
intr-o populrrtie bacterian[
pufine brctcr ii sunt in stare

corlrpclc|'i

TIANZL:O i'

E 5-10-15 Irrilrte
E controlali rciic

'j

".-"'

in perioada

d. Jcz\oltare a

populaliei brctcriene,
ls alte bacttrii

de

LJ

()
.--- *.-/ (
-:.,*..r'
:,

demonstrati la streptococi

Ill

Cmrt

jicmr*!*
sts|{

\- @l I
la-*
I '.:ilt-5o

(;b;
t
r-;."ii
\,.=*_--l,

[ '"**l

[l)rwts{.[d

Transformarea naturala la bacterii Gram pozitive

t)'il,
at:'flttir.i

'

r: TlTli[

$r,iiriitr*.*,

-_-riNffi rc
{{)i4}r

,ri1r*ia

05.L2.2012

Nehserla gonorrhoeae

hiermmm
r;:;',,cf9mo

cc;1.

:\
i

carl

/l"" rgnbrsa

. de zeci de ori ln organismul


uman,
. evolulia cronictr a gonoreei
. inainte de descoperirea
antibioticelor gi infoducerea
antibioticoterapiei,
. gonoreea - ani de zile

i
e
I

Sldntlor 0,rA !nrero.Nr'obthiS grndk.oqd.K


h a dt
r5i4N@sr$@d"tu!
h*,ls 1D,.ier!i r.. r. ri..,..',.1 r.'qse*lerrir+rlr
si ilrf.rr:; r:r:l
$trr r{rf rir$lfr: j r,.. 1 rrr.r...?r! ) i' hlad!&aans ri*rq nuil&bnwtdby{x
.,:JtlrMalN.i;?:,?tai
s\nr,r\.ij i'rF$a r : .. : r
rr:,,
o +.
:61{g.s*hrr.:r3,@sryG.m|@

f&r$m gaatrioeft

r2/5/2at2

h&

N, gorarrhoeae

Translbrmrre n:rturala la bacterii Gram negative

El
Ir ,*r

L2,51,0r2

,.-.. -

]
-;..-:-

Ttansformasoml:
vesicule ,

tr
tr
E

^iJ
' r",c

Nonlo t$1.

.!'

::j:\

ADN d.c degadat


ADNm.e
Recombinarrcou
cromozomul bactR

:; . {
"..;.' i.
Pl&
lodg|hN.

I i.illural al transformilrii

:,'rirl genetic

PFfor

t2i'i2ol2

Xonlo runl.

Neisseria gonorrhoeae - gonococci

t
E Structura pililor
dererminattr genetic, ']-t;
E::::i:_Fl-d

lJ

ln crOmoz0mul OaCIenAn
gene pentru pileirl

a ,,

,g

,
n i"-r.,i",irl"".**,
i. -.-^-,r d^n^^Mir^, r'u:':+:'t$'.
,i'' ,'i.
o
i,, j,.#h,,,
E gcnaactivtr

.,*"'Wr4it|,
ffui:i '

o',r'Jil.T:X,|"""0''
r

neexprimatefenotipic,
codificd alt[ structurf, a

,1,

"i

sr|t,t,,,*

pileinei

05.t2;20L2

Gonococii liza(i,

prin ant corpii fomafi gi

elibereazi

E
E
E
E

Ngmcntc de

AI)N,

ir

pdtrundc

se recornl i,

de

i
&

contjn ! jne tdcute pentru

pilcirri
Fragmcnlr:l

;@l
Wl

',p
\s\
rB

.rcFffitffi

t8$swd*

,11;?,::r-.'+r.

ll

aa

a"rfr*

tr:D't reeCPJs)

ADN exogen

:onococii nelizafi,

cu gena pentru

prlciili,

E
E

rtc,,trr

rar c

orrr.loagd

o noLra rc

lLlrtclr i:ltl11,

0 [i)r i :lructufe.
iu;riir,i,ei-

t-O(liii
-

tr

",^

Pilii induc anticorpi specifici, cu rol protertor

E Anticorpii fomnfi nntetinr qmt in4cfivi n$Itr

nceslnr

gonococi, dar se fonneazl alfi anticorpi 9i fenomenul se repettr

-artl:.

4i Fansfert pni fansformation


S

-:A .:

t.,l'r r:i i1r,rai,:'r,!*,'rg

pNnh.4.

^1

eii*.

,,

u.d,ra
'7s \

Feptocoques'v iiida n$'

^S.3

$:

o\*f

;
E*

"'*

ADil

libir.

pneumocoque

Repararc
2.Atatare4 integrarea in Gonococi
nelizali

Fr

tl

4.,
i:, Xi 3l i3 *: i
!tttliri

ir ,'i'j
' llN liber cu
:Jlla tacutd
I inactiv[) pt pili

; w: kr*

31

exprimarea genei
tacute (ex. P2)

05.L2.20L2

Inrportan{a experimentalI a transformilrii


transformarea artificialS
tr.*o*liaa*i"!+lakaaq*+fb
plasrnidc,

pernite araliza geneticd

lntocmirea hf,4ilor

fini,

p-

genetrce,

tehnolo.ix ADN-ului

lJ

ln!ilrcl,.r !rllJilcl

recorrt)if..rt

w@
v

Sr

.i

..

3
@@
s{q*

tra nsformarea

artilicialii

E trrrq"'r de ADN plasmidic sau u.,,..,,'",:,,',.-r...


r -+ J:-li ":
wi-..1-' ., r.ci;rri;;i.;;:!*ry
E n'r .,r r . :.r. .ic c'r rl'elentA. ;." 1""
EI crLltivrrle r^tr prczenta unei cantitdli .r '': t
crescute de CaCl2,
ii.'rqn ..n:*
E produ:e rrrici brc$e in peretele t::lt',."r=-","u]
: | {h!
c(l..li I l:i l)aclcilcl facciltoare
tr S, pr .lrr: Ju:'t 'r'r " .' -' elrPe:
E alr5rrt,,rcrttrirrr, iiri
ilrl.,:.rE l rf.l
citr,;

ELIa

4':
' t l

':::::

;:i:

:::.':I

de
/i"*i

i*d*,
..*\-r-*-l

i;,rr

10

05.L2.20L2

moleculeindependenle,

f'l

qrtnnnme in ni1^ntecma h'.tpn'n,

lnserate in cromozom

Material genetic ertra cromozomial

Eplasmide F

E rcpliconi,
Prof' Lia Monica Junie
E autoreplicare
ru.lzoLz
** "-il
"nl----5G612012--__i

tr
E

molecule ADN

dublucatenare,

Cl circulare

El

|J

Covalente

bene

Ba{brialc*ll

PI.ASMIDE
Bacterial

DNA

Plasnrids

grurs

'

E proprietati bacteriane,
E neimportante ptr multiplicare

iilflrfi llr*l ihililnt*

ttt* t:l:: x \l+t1 r it.l

it

bactenan'f,i:!,

05.L2.20L2

replicares plasmidelor: diviziune bacteriana

SftndErcutromo-enrd
hacterian-----------

/TT'T|-\
replicare

bidirectionala

/1-TTi-\

tss t t8s
I t'Xl lo\Fl
l

model O

t.3ej

E3

rea bidirec{ionall a plasmidelor in cursul


diviziunii bacteriane
Transferulplasmi{g161 verticailL la

#)
,.J.l,i,lll|on o'"ji*'""
t

metaborice

OP'ne cdtabolisn

&iinolreplsatts

molecule ADN auto -replioative

Plasmide

gene de incompatibilitate: grupe

cdl cycls o( one pbwd

cgll cyctc d 6cry5tbb

Fletrsd

@e@qe

d@e-

ffw O ) auacelEi
dO O?
r locdeata5arep**"''ffior.*o*
ffiL'*nff

1,i!.1.

'ffi

c*n

!fi'J,xl "nr,"."

F.H,!{,ijiH-

*;u* l:

ffi',ffiffiffi
ce(h tglt hrs cog d t{fr

ptt6rrri'd

trr(:off!,dil)l*' Dbarffflf' lixtr tsatr


u,glrrwreo lo anns,lr usr.}

Dintr- un gruD de incomDatibilitate:


diferite
pot@Siista tnt-o bacteriet'dqft'pbtt0.exista lnf-o bacterie 12
din grupe

05.12 .20L2

Transferul plasmidelor

feromoni

C*tasl1*:sffi*rl

Fleromsnes in

llants

h
ri

!.
tr Conjugare
tr la alte bacterii din
r1 aceea;i

tr

o,f.,*

plasmide conJugante

.
.

specie,

specii sau genuri diferite,

auto -transferabil,
autonome in citoplasma

bacteriilor,
rntesraE rn cromozom

Fheromones in Animals

";::,r"'' \

,:;::,.w1

ffi'*
;;:;.,,,;---

hansfer de material genetic


Eintre bacterii donatoare (D)

9i receptoare (R)

FEROMONI - biosensori in natura

Insecte

' .:
^

contact cetular direct bacterian

,rl

llplasmidic,

Elcromozomial

CONJUGARE BACTERIANA

Conjugarea la bacterii Gram (+)

transferADN:

pili de sex Bacteria

05.12.20L2

Bscteriadonatoare-D

tr

sintetizeazi gi exprimll la

proteine de

I
I

stassre,

de aderare,

"aggregationsubstance"
(AS),

n
n

sta$re de bacteria R

tr

molerula de plasmid.il

bacterie D masculintr
F+, R+ or col+

2 bacterii alipite

transferat.l din bacteria D ln


bacteria R"

r
I
r

alungite.

F, R,col

Pil de sex
ata;arecelulaR,
bactrii feminine R

t
I
I
I
I

nu plasmide,
F, R-col'
sculteJ groase

receptorilorproteici
membrm eltemd
zone de contact pe
suprafala celulei R

t*#;;
I

E
B

' :::t

Perechi bacterii
F+

F- ;Hrf+ - F-

U KT-K.

ro.E rOOUu*Col-

feromoni de

operon

sex

"Tra" - RTF (regiune factor transfer)

@
E
tr
E
E
E
tr
tr
tr

produce feromonide sex

(peptidecuGMmic[)
Efect sfrodisiac asupre becteriel D
dob8ndireaplasmidei,
numai produce feromoni de sex
producealfi feromonide soq
activi asupraaltorbacterii,

caredefinalttipdeplasmide

lvslzo(2

tuf@r tonlq lu.l.

20

CONJUGAREA PRIN PILI DE SEX


alipirea bacteriilor

rtjdtxlwffildftrbitu,lxtffi
rdolpd!{r?dFa{i*ttrf@B{
r brege ln peretele bacteriei R

punte citoplasmatica intercelulari

fih+ii4;ti.*
hgSxs*#l):I.
c

coli, bacil piocianic, etc.)

liwttlsrtFialh*4tr,r+t':
il{n*41{*ir@m4rtcr4*

*P{a}#tu*fi

I lltt!:b'}9}*wfi&l
hra.:irtiaEi4j$9.

+ttr4F{

bacteria receptoare

05.L2.20L2

plasmidele R - structura

transfer plasmidr

't"7=*r(iir.orylgryrur*r- l
\."* *y' 'l--*r.ifSr*' .i'*-*-*
I

a-kt

btxq&F

d'&tu d *i d ffi
-plasmidaff"*
.qin"e_:*_. li
li
//.\-T--.':!
\ \'rqk'r,*j/ .) ji {i nem4iepit lli!

td""

tr
tr
tr

tr cetevagenc- r

*;.:;'m g;* H-'n::,


* .d :"y"',".':x:*e:uix
rrlti
I
endonucleaze'
fil
rl***x
sec[oneazaocatenlamoleculeide

\,

.)

3 - 7 gene,,r"

i f i'

i...t i.::t 6p11 pl65midis


ut..h.,*rdhrd

b6et.*6qi

sute de gene

Operonul Tra gi RFT identice,

unidirectionaltr,

Plasmide R

E
tr

transmitepidemic
in populaliile bacteriene

tr
r
I
I

pe orizontalt,

se

tr#
I

inl

*o6"*"*kttu|ffiFb&*!Pei

cltoplrEmrtic

i
j
I
l.**.*w

Crtenrnourintetlzstl
- tece prin puntea citoplasmaticl

Uacterian"tcatevaminute

@*..',*F

B.tF6t..[

*..
^\"ffi{i : r/il
t l;\-.--..+/ }
\-.*r':ii
.J
v

w.wr-'.**rs
".ohfiasawmErffild

@*q

'rsd

ra*H/a

Replicare in R
- NIatriP sinteza catenei

complementare
-repticaze
otcmd. * oE dd.rr.,nr
*r**{

'ffiffi-

"ffitW
',Wz
.WM
*"'*\

ln

ladescendenfi

conju8iarebacteriani

tJjei*rt*1*9#Y.?

bacteriidin
speciidiferitedelabacil
coli la bacil dizenteric
genuri diferite

\'-,

60 plasmide gene
rezistsnta la p lactamine

rezistentala md multe mtibiofice (3-

ht*1qD)ot!'r.

E
E

Genele

rr" =transpozon!

plasmideR auprovenit din


plasmide F,

|etuFtr

cromozomul bacterian

05.L2.20L2

conjug&rea R+ - Rlh.d

ffiffi--k @
//ts"*

HH

***

rr..:t::i::::r::i.:.:,:::l:ri:: -i,.

M.

*- -

bacteriadonatoareR+,

k4Jffnt

bacteriereceptoareR-

bacteriaR- devinR+

plasmide - gene ^rtructurale


proprietati bacteriane

Conjugrrea R+-R-

El

mecanismulprimordialde

labaotriile Gramnegativc

tr
tr

r
I

rezistenFconcomitenti

gene de rezistenla

bacteriilerezistilla

oonfer[ bacteriilor
dezinfecfie

antiseptice,

slruri

de metale

gele

r Pb,Hg,Agarseq
E detergenfi,
tr radiafii,
E bacteriofagi,
tr Colicine

la 3-7 AB,

E rezistentahbloclaAB
tr Bacteriimultiplurezistente
E se selecteazdln
B mediul spitalicesc
tr infecfii intraspitalicegti grave,
uneori letale

r!f*47

6itPl'F,!+.o$!

i- ftr,fr:Kil'1\ astaa ;

- ,rytbnD
;,\'Lr!tftq!J
\ '"dL {'/
., )6a!(il?"i
r

cre

;',:{'""
lXT+;\'"4*x
,
*lr*a)
,aicl5r,f
Rl4#1 '
ji

Mcanisme de nsistents
le antibiotice

a. nOtO
t rd*&lt

b' R'$t'i

uavar'

Plasmidele

gene pentru sinteza de

tr
tr

dtebacterii:

Jgli$Eglabaoilulcoli,

\
$$ct.nffi Rc(sMry
hcldb.ln
bddl"{
Dr ks
plasmide R fara operon Tr4 non- traasferabile

G.w

Nuconjugare,

transmisela alte bactsrii:

- o

tur

ionto

lut.

33

>>

Stafilococ . rtsfilocinc

bacil piocianio
(Pseudomonas) -

I{ebsiella - klebocine tulnini de bacil coli

piocine;

rClostridiumperfringens

Transformare

Lust2oiz----

col

- perfringocine

xeti$larFe

,A,W
<<

I
r

omoaraprincolicine,
tulpini patogene de bacil coli

E bacteriocine
-:-'--.:--spectrurcstrgnsdeacliune
u
= pr8|rmloe
bacteriocinogene I aceeEi sPecie
*r.fffiFfiTffiffidft
Lur,t2

05.t2.20L2

,,.*u****,,9t.doclne! rntagonism baglerian

conjugfiercol+-col-

(o)
/\

bacteriaD

cal*

tr

bacterie Rcor"

E
tr

#-^.
.i",,,{

)f tr

Plssmidescol' - Becteriocinogene

Wb

t
u,.

c:7il

E*";, -:* ,#*U. l, ,

\_-/
Bacteriacol- devine bacterie
col+,
Producebacteriocine

seamplificlproprietatea
bacteriilor de

produce

. faringiani

Bacteriocine

plasmide bactericinoge,ne

5i$r]Sit:v*l
ssuprr f\.-dryx*
E@h\e3
rltorbacterii -.J
efect letsl

(iir rhe
{)*i tx}t rncr,r
I
\
thr ro;r ol
lcft.i.t*c /
\
,,.rar, ,/
\ r*:,;tic,."'
Omolecul[debacteriocind \ a
//
omoar[o bacterie
Rf 51{tn1!r

il
?t:}

l}n{:tsri{xirr a*<l a:{r*.r


{r:* lsriir thp r$ef f)l

-i

'.,.."

" "/

Rr$lsf*f}t

pf$*l]{Ii$.r}
Bacteriile producltosrc de bacteriocine
- imune la bacteriocinele secretate

N*turr Rwiervr ] irticrr:bialcgy

patogenitate/virulenta

uro

r
a

lezars

Lt titonlismatice

Psrogen

epiteliu colonio uman


Bacildiznleric

(:..* *
pertubaCneigizareal " "=
Dermeab illlatealnefi ibranei

E. coli enteroinvaziv @IEC)

"

05.L2.20L2

patogenitrte/virulenta

Gene
l-l "rili-..crrkcsot nrr#tirr
catabolizarlactoza

i
s.,.1

H-

l.
;S
, Ir*'i@
I

r
I

-r ,l '
l{

Irrh6l,vq
M.lqls,*3q
t.fu.f
ddMr
lldffl!'lfu$,
t8shnwd
lrlMr@+*re
ffW'M.

Salmonella,

produceaHrS

El

E, coli,
producere energie/substrat
energetic

a
r

Pseudomonas:

toxinebacteriene . plasmidele se pot dob6ndi sau se pot


I Stafilococi: toxina pierde,
exfoliativa, . variabititatea patogenittr ii unor
rto
ffn.BemolizinaE cor- hbr-iliiiii)iih
43

Producerea de tumori la plante

Transpozoni & plasmide


entrotoxina vibrion holeric

Plasmida codifica

_+_{
LU5/20L2

a,

P.ol@r No.to J!ni.

Plasmidele F - de fertilitate

E molecule de ADN d.c,


E circulare,

trffi
tr
E
E

GM 1,4 x 106 Dal.


35 x 103 perechi de

trt

baze

FPlmlrl
1$0 kb

plasmide criptice:

Nu codifica caracter

transfer material genetic

bacterian

I
I

vibrioncholeri

1""i.3

?6

StrUCtUra

mecanismde actiune,

Similare
E coli enterotoxisen sindrom
diareic

LU5aOL2

hbr

ionlo )unl.

&
?: irsrdaflri i+q'rerrrr$
'S
Tn1*Ct
= !$nsF*son

= *idir 0f !rpii*ilir.r
tll = o!8* {t r{rrltnl lr?nt{9r
lrA s lrn nJdctohi
Eat1

45

,ustzolz

p.r.-, ronr- ru*ii scoYclrl9 u9 ursclF9 4

05.L2.2012

Plasmide F - structura

o operon "Tta"

(de transfer),

gene de fertilitate,

E complexgene
(2r),
tr fra$fer

-IS: secvenfe de inserfie,


- plasmidn

material genetic
prin conjugarc

- cromozombacterian

Eformarea pilului
de sx

plasmide conjugante

crom06larhsterlin

F Fl4rnid-.m.,1,

desooperitein

,p-ffi**r{l_ffi$
tqffi*FSry*X*&
ffi?t$*;t*
.s'cdL'-.trl1. r*-o:dr'\:._}*"ltr.qel'.Y*1rd5'

fenomenul de
conjugare bacteriantr

ln populafia bacteriml

l*

r*,.S"{}
^"1..*

..

lJ EansmlteremlecEoBsa
--!F-r

t*.r.

..4.-

EeKlrt

E peverticell
! la descendenfi

tr

cir(ular,.rtochromctofril OilA mol.csl.

grctwid chrwrrro*o

$tfr'!lo|W
{c}

Cqri{ldis

.nd |trn*ls

d.{ f

pb$mid lmm

Fr dner to rn

f- reipifit

peorizontaltr,

conjugare bacterisni

specificitate de

la bacterii din acea$i

"" r*:
r.*lr i-lUi

:"\.nJ,
*"' rllrcdld

,^Ki-

specie
{bl

.
.

specle

R.pLfild trfier

!ffir

tof

iltiblotit rasislam

intro

populafie debacteriiF+
2 sttrri alternative,

malorl,Ji{iZ dmcelulele !: devmbf

Conjugarea f,'+ - F.Ata$arc bacterii

. Dunte de lesilturl
. ieplicarea ife fansfer

r*E{d

fiw

dilss
*t

hlS#i x fhdq&rtondh

uEftsibfirerfl

et

tur

N?*4 t"t **

f ilf ,rwrtd

hifdealnizarea

&s

$*

baoteriilor F-

05.L2.20L2

Gene de metabolism ADN, gene de interacfiune cu


alte molecule de ADN

Intcgrarer plasmidel

tr'

in cromozomul bacterian

i"-}

.i-l
il*"""-)
: 1", i
.\i*iAi .-:,.. 1i

tr alte plasmide
E integrareagiexcizia
pnsmloelor
E ?ncromozomul
bacterian Si
E cointesarea

{"'i I

\Ji lv, .v;

plasmidelor

lltr

Insertia plasmidei tr'in cromozom


E coll
Scvnte

omoloage

Secvenle de insrtie IS

tr

bacterie

tr

celule masculine (supermasouline)


o bacterie din lOsbacterii F+

Efr (high frewency of recombination)

integrarea plasmidei in cromozomul bacterian

. Recombinare omoloaga ,

Identice pe
cromozom
Elemente

> 90%dinADN-ul
Df, ,glpgomoloage pe croqnnergurl,bacfriar/pe

plasmidl

Insertia plasmidei F in cromozom


prin crossing-over
;f,

colJ

origine (O) insertie


- insertia plasmid F
- gene a, d, c, b cromozomiale
- rntegnzl

E
^
u

pl-ahidsF iategrtr i8 crooozomul


LUal20L2

.- ^

InarKen

l.z

ffir

haclrian-

bniq luii.

10

05.12.20L2

$ADN ..onro"omial E. coli

{-

_}

?6i-:i'doiii$amulul.lprcrian,
unidfuecfional!, modelul inelului rotativ (o) in 100 sdnut,
Transferul de material cronozomial de la bacterie Hfr donaloarc,la
neplicuea

bacteria F- receptoare, depilde de durata contactului dintre cele 2


bacterii.
Rar se poate taosfra lntreaga catnll complementarlaADN-ului

cromommiaffie#
rudUfCltlie rarian in contactPf00'rf,fl111181'

Conjugarea intrc o bacterie


,'{igh

Gene cromozomiale codifica oroDrietati bacteriane : biosinteza


arninoacizi, degrldare zqhar, iezistarfa artibiotic, sinreza pili
L2/s/2o12
Prot .or lonlc. lunl.
&

Bacteriile Hfr pot transfera material genetic cromozomial,


prin conjugare bacteriantr celulelor F-

si bacteria receptoarc F-

Frtqrtruy Rcsan}binaa! uXrh;l$ ;*n

hs$l t$

el,r+s;ffiisi*! }llA

d!){i.l1{

6'*ipifill {r!l

Cwofhoildtofibrofir$ilh
inta$dotl F.:mmain3 in donot csll

E
tr

IIfr

51

Rscsmbffiuffi
bciiffsend((loriltd

'.}

Hfr

Rrcsmbindrl

.*i!r*i

tPtilxs{tt

localizirea

Brcteriile Hfr (high ti'ecvency of recombination)

lnaintea

pot transfera material genetic cromosomial la celule F- prin


co'yugare bacte'ial^
?rbr ionlo lunr.
lu5l4l2

Transfer de gene cromozomiale

llk

\r

52

bscteriadonatoareHfrt

I.

bacteri{rcceptoareF-

Jt

+",1\i7:
k!l!&!.i!si*r*!
hi6rksws*
"xlSVdrFaktq*

brcterh rcceptorr F- diferite gene cromommiale in


functie de localizarea plasmidei in cromozomul baoteriei donatoaro
Se transfera la

Hfr

Bacteric R - tr'Jrimegte gene cromozomiale, nu


hb' Nonrc.
rdlnlltetecterie

F-

geneid

treclnbacteriareceptoare
genele A, B, C, D
ultima trece olasmida

li

,ti.-.11'

\\t

I
r
I

lt!tr

1l
It

runi.

/-\
'&,2- \-9I*g'^L-*,)'
z\

tr

Si plaqmida4

@
@
R

plasmida F integata in diferite situsuri in DNA cromozomul

5!

11

05.r2.20t2

gene

cromozomiale.

transferateinbacteriaR

E coli - tuloini suxotrofe

iI

+,*l

rccombination)
r-1

recombinare omoloagtr

n
n

rccombinant:
l-2 pmprietlfi ln plus

l.

2.
3.

biosintezaaminoacizi.
degradwezahal
rezistantaantibiotic.

Pil

arsFrai{ll
ttdk'rlb*dCu

I ftt$tfidi

Hrb{|$ff

,t{rJmftlH tffdt

,il ::::1:1

trrhri$S.

I "_-,.-

[s ctrd{tirt}Jrrk.re h I lkhwl}dtt{ffil
t$l|q'lffilihlR}s
dlkts*l${ttr4it
(Oistts,$ldkifttr1, wll$['l8,esl

t
'

r,,,.,,,r:r:. ." ."

.i:"
recombinant prctotrof

ilef*bio*thr* 1c-,-70

phe*cyst

Experimenl Lederberg and Tatum

recombinare omoloaga

recombitraree

bscteriilor' , '

*"*s*qx-gtryf,e*

**^i1*ia;.

Rcombinstion
llomolggour RGombinstion
lloqologour
(HF|l
s\$Kn@k.ldFiiNn.ilijliiillj@PrclGor
lUSnOtZ

: : : :.......;:....:.a=:...:=

ilonic.

k:'IU.a{dFMtn,$*
e

d*

runi.

Transfer de gene cromozomiale

68

Conjugarea intrerupttr - Wollmann

Expenment Lederberg md Tatum


ta?it

}ii1

'$fi

bscterii Efr prototrofe: capabile de a-Si sinteti?. singure toli


constituienfii celulari 9i sensibile la StreptomicinArt "
bacterii F- poliauxotrofe: mutatii nu pot sintetiza un numlr mare de
constituienfi (markei genetici lac, gaD 9i
ffi&Mt{&

t2/5/20t2

Prof.sor Monic.

stf

iuni.

t2

12

05.t2.20r2

Serductis - Trsnsfer de materisl plasmidic


prin factor de ser

Ai

cromommial

.,,.,_,..,....,,jt.:r,..,....-.1
:. ..
Dplasmida F;': conline gene cromozomiale pedtiii uii cdifia;
bacterian: genaA
O poate fi tusferatl bacteriei R
tr se poate inrcgra ln cromozomul bacteria& sau

;o,;.-.1"--: ':::: ', , ':::...,::::::::: it"il1:.;.:-.i:::.::::;:iffllliilii\\[iii,l't\iW


> s-a lntrerupt conjugarea prin agitare, la intewale variabile
) s-a cultivat bacteria recombinantd oe medii de culrurL
D pentru a observa ce proprietS!i au iost r,,nsferale
> s-e+tabilr+leedizareai.X:"Lo,:,ffi":
bacterian"'ironrul
t

harta geneticl a cromozonru]Lii bacterian la E. coli.


\: sll:l r;lrc's d tlx alaorm df.

Plasmidele

gt

i
I rr*ndhsil ltrWrpm
ithnl'$rl1
i
\ *eied ldir I

E
E

!
\ riturrtiok!) //
,\
t-.
-/
I )rnrsmNn \
I
lt't't'n
!u-.*-.*'.-

cromozomi de sex
gene pentru

proprietifi

bactedene

E
,*

lscttse.
Iiis crll- con use lsctrse.

E 400Ogeneoromozom
ffi
1ffi1:'*
h&r

bacteriile
minicromozomisau
cromommi faculativi

E5*otoq*:m

pot integra sau nu in


cromozomul bacterian sau
ln alte plasmide
Se

La alte bacterii, hartageneticd


u

ilon,Lr

s I

lJn.

e )ad ebine,ethoseufl

Molecule

tr

bacteriile le pot c4tiga sau


pierde,

El circuld intre tulpinile


E unei specii bacteriene,
El intrespeoiigi genuridiferite

tu5/2orz

7a

excizia plasmidei F din cronrozontul bacterian

corecttr,

plasmida se desprinde

ln tolalitate
tece in citoplasmd,

plrsmidllautonomi

lncorecttr:

Ioparteaplasmidei
rdm6ne

-I

ln

!NIc(;ti r t!rl

()-'3'-*
IXClSlONi!

l$ tilf l$t{

.ri

cromozomul bacterian
I
i
i,
",...:...q{
plasmida preia gene
r 000bac1eriiV
intr-o b,r.5;;. ,1i
oln cromozomu
'
o bacter,e rli,r | 0 - I 15 bacterii Hfr
bacterian
I Jii J(\ ene F+
+ plasmida
: hi5ledfr

r+

t'

F':

plasmid[hibridd,

i'1,
\*

/t
i,
\J
l1

13