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BIOOIA TN MIKPOOPANIMN ANEITHMIAKE EKOEI KPHTH
, , (( ) ) ..
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((screening) screening) (( ))
(( DNA,DNA, ) ) ((, , , , ))
((point mutations)point mutations)
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: : (( ) ) (( DNA).DNA).
(( ))
DNA DNA 1010--77--
1010--1111 . . 1000 1000 , , 1010--44 1010--88 . . 10108 8 101099 ml ml
. . ((hot hot spots)spots) DNADNA
To RNATo RNA DNADNA
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DNADNA (( SOS)SOS)
Ames
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(( ) ) . . E. coli E. coli (( 25 25 ))
RecBCD, RecA, SSB
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DNA - . DNAGram- : DNAGram+ : DNA
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((R)R)
GramGram-- . . . . E. coli E. coli , , Pseudomonas Pseudomonas sppspp.. ..
15 15 , , . .
4 4 . . . . . .
--
: : 2 2
FF FF
(( ) ) (( ) ) ..
tratra FF
tratra ((sex sex piluspilus))
((RNARNA )) F.F.
DNA
HfrHfr
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FF HfrHfr (High Frequency of recombination). (High Frequency of recombination). FF FF++ ((--) ) FF FF-- (-).
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Hfr. -
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3 3 :: (IS)(IS), , ((TransposonsTransposons)) ..
: : DNADNA
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(IS2) (Tn5)
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(Integrons): . (). . . .
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(( )) ((Southern blot, Northern blot)Southern blot, Northern blot) DNADNA ((,,, ,
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DNA DNA . . 60 60
DNADNA (( 44--8 8 )) . .
(( ). ). DNADNA ((sticky)sticky) ((blundblund))
((isoschizomersisoschizomers).). ! !
DNA DNA ??
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(( ) ) DNA DNA DNADNA
EcoRI Escherichia coli 5 .
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() (DNA-DNA DNA-RNA) ( 32) . , .
() DNA in vitro DNA DNA . Sanger DNA
20-100 in vitro( ) .
: H: H
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((, , , , )) (( )) In vitro In vitro (( , ,
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1 2 DNA (40-45 kb) ( ) DNA.
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1. DNA2. DNA ( , 3. ()4. ( )
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DNADNA ((DH5DH5, , 101, 101,
JM109)JM109) ..
CaClCaCl22 RbClRbCl
DNA DNA . .
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(msec) DNA: :
, PTG (), X-gal ( ). . (satellite colonies). ? ?
((PCR)PCR)
in vivoin vivo . . DNADNA in vitroin vitro (( DNADNA 101099))
PCR: PCR: DNA DNA (( ). ). 4 4 ((dATPdATP, , dTTPdTTP, , dGTPdGTP, , dCTPdCTP), ), ((TaqTaq, , PfuPfu),), MgMg , , (( 1818--21 21 ) )
PCRA) DNADNA--) ) ) ) DNADNA DNADNA DNADNA--
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PCRPCR , ,
PCRPCR ((RTRT--
PCR)PCR) ((, ,
))
(site-directed mutagenesis)
- - - - Hfr : in vitro DNA ? : H ? () () (PCR)- PCR