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''

.. , .
DNA RNA 4 .
:
1. , :
. RNA DNA
* () (G).
:
1) (C),
2) (U)
3) (T).

RNA C U, DNA C

U T.

.. ,

. DNA
5% C 5 , DNA
5-,
. RNA
.
, -- -, , ,
. ,
..
2. , -D- ..
( RNA ) -D- ..
( DNA ).

--,

-9 -1 .

( ) ,

(H3PO4)

3-5 (

).

.. 5

3 .

..

) :
1. RNA -- AUGC
2. DNA -- dAdTdGdC(d ).
) .
1. .
2. .
3. .


.. :

1. ..
..
DNA
:
1.

3---C-T-A-G---5
5---G-A-T-C---3

2. Satellite ( DNA )
.
DNA.
3---A-T-A-T-A-T-A-T-A-T---5
3---G-C-G-C-G-C-G-C-G-C---5


( Watson & Crick )

To 1953 o James Watson Francis Crick (W. & C.)

DNA.

. W. & C.
Rosalind Franklin & Maurice Wilkins DNA (

) , Erwin Chargaff
.

.Chargaff (1950) :
1. , GC.
G C ( ).
2. A+G = C+T.

:
1. DNA
, .
2.
.
.
3. 23,7A. 3,4A.
() 10,4
34,5A.

4.
2 G C .
5.
. .

. W & C G
C, .
,
(
).

W & C -.

( 75%) - )
11 )
20 ) .

DNA G
C . 12 .
, .
,
- . DNA.

RNA
DNA, RNA
.

. ( RNA )
- DNA A-RNA RNA-11
11 , 30A ,
14 .

RNA-DNA -.

RNA DNA


RNA .

RNA

:
1. tRNA ( RNA ).

10-20% ..

25-30 kDa ( 70 80 ).

( ).

2. rRNA ( RNA 1/2 2/3


).

80% ..

rRNA
23S ( 1,2 x 103 kd 3700
),16S ( 0,55x 103 kd - 1700 ) 5S ( 360 kd 120
). 1S=10-13 sec.

: 28S, 5,8S 5S

18S .

3. mRNA ( RNA
).
5% ..
500 2500 kd.

, Drosophila 4, 40

1000 E.Coli.
.

DNA.

To DNA ,

histones ( )
.
.
:
1. - ( )
2. - ( ).
(1)

DNA.
(2) DNA 0,5 NaCl .
(3) 5 1, 2, 2, 3 4.
(4)
1 4 Lys Arg.
1 4


,
.
(5) :

,
DNA .


Roger Kornberg
1974 :
1. 2, 2, 3 4 1.
2. 100A.
3.
100A
( ds ) DNA.
4. dsDNA
, DNA
.
200 bp
dsDNA( base pairs ).
5.

(3)2(4)2 .
R. Kornberg :

() (2)2(2)2(3)2(4)2
200 bp DNA.

( linger ).

DNA ( Core )

1(

). 1
nucleosome core practicle ( ),
146 bp DNA .

DNA

linger DNA ( DNA ),

8 114 bp
, 55 bp.

70A 110A. (3)2(4)2

2-2
.

DNA 1

166 bp.

1 DNA

DNA
DNA
.

166 bp DNA 10 ( 560A


56A ).
-

1 .
300A .

Aaron Klug : 300A

100A 6
110A ( ).
1
( C
).
40.

-
scaffold ,
.

DNA

.
15 30 m ( 45 90 kb)
300A 0,6 m.

.
0,3 m 0,4 m scaffold
1 m.

140.000.000 bp

2.000 70 kb. 0,4 m scaffold


6 m ,
.

300A

non-histone proteins (- )

.
.

( Denaturation &

Renaturation)
:
1. pH 6 7,2
2. 30 40 C
3. - 0,1 )

DNA RNA .

DNA ,
. DNA
.

( Denaturation ) DNA

DNA, DNA
DNA
.

melting ,

( melting temperature ) Tm

DNA .

DNA

260nm ( UV )
. 40%.

DNA.

Tm DNA . DNA

G-C Tm , - .
m DNA
G-C 77 - 100 C G-C
20 78% G-C ( )
- ( ).

.
Renaturation ( ).
.

, DNA ( 94 C )

4 C

. .

pH.

..

W & C
.. pH 2 pH 12
DNA.

.. 4
:
1. ( Solution methods )
2.
3.
4..

.. .

.. .
/ CHCl3
.

..


.. .

.. DNA & RNA

.
RNA DNA-
DNA RNA-. DNA RNA
.

..

EDTA
.


..

DNA

Ca3(PO4)2.

DNA (dsDNA)
. Ca3(PO4)2
dsDNA ,
RNA ,
( /) DNA.

DNA (ssDNA)
/ dsDNA.
DNA
.

Ca3(PO4)2 DNA ssDNA


/ ,
dsDNA ssDNA
.

Tm dsDNA G-C

dsDNA
.

(affinity chromatography)

.. .. mRNA
( A ) 3 ( poly A tale ). mRNA

poly(U) (T) . poly(A)
poly(U)-(T)
.


.
.

PAGE (polyacrylamide gel electrophoresis )

DNA RNA ,

bp.
,
DNA ( DNA sequencing ).

100.000

bp. (%)
DNA, ..
DNA.

CsCl (equilibrium density gradient ultracentifugation)


DNA.

() CsCl
.

p (buoyant density) ds-Cs+-DNA

(P=1.660 + 0.098* XG+C) DNA


.

ssDNA dsDNA

RNA CsCl Cs2SO4.

T RNA-DNA CsCl

dsDNA.

(72 ).

DNA

DNA .
( DNA),
.
-.

DNA

.
. DNA
.

:
1. DNA
. DNA
.
2.
.


(

.

H :

L=T+W
:
L Linking number DNA
.
T Twisting number
DNA .

W Writhing number
.

W , L
.

DNA = 23 W = 0 , DNA
= 25 W = -2 . DNA
DNA L = 23.

L ( W)
,
.

DNA
.

DNA
0.03 0.09. DNA
.
.

DNA
.

DNA
,
.

DNA ,
,
.

DNA.

. DNA
:
1. DNA,

2. DNA
3. DNA.

DNA.

(ATP) .
(Tyr),
,
. 5
.
- 3
.

(DNA )

DNA ATP.

200 bp DNA .

, ATP DNA.
DNA
.
. DNA
,
.

DNA ,

DNA .
.

DNA DNA

DNA

DNA

.
.

DNA . ,

,
DNA.
ATP DNA .

DNA

1955 . Kornberg . Coli


DNA.

DNA

(DNA-polymerase)

(dNTP)
dNTP
:
dNTP + (dNMP)n (dNMP)n+1 + PPi

DNA .

DNA :

dNTPs
.
G C .

DNA .
5 3 (5
3).

DNA.
W & C DNA
, DNA.
.

1. DNA- 103 kd
.

3 5 .

DNA. To
3 DNA.
3- ,
. DNA
,
-, 3 5

,
.

3 5
DNA,
.
. ,
.

5 3 .

DNA 5 .

5 3
3 5:
(1)

.
(2)

5
.
(3)

5 3

DNA. ,
3 5 .

5 3 -

DNA, DNA .

5 3 3 5

. , 5 3


DNA .

2. DNA DNA
& DNA pol. I
:
1. - DNA dNTPs.
2. DNA
3-
3. 5 3.
4. 3 5 .
:

DNA pol. III


DNA, DNA pol. I
.

DNA pol. II DNA


DNA.

DNA pol. III

10 900 kd.

DNA

pol.

III

, -
109 1010
.


, 20 DNA pol. I.


10/sec DNA pol. I.

DNA pol. III :



DNA.

.
, 3 5
, 2

.
DNA

.
dsDNA.

1. ( dnaB protein)
. 300 kd 20
(E. coli).
2. ( Primase )
RNA (primers). 60 kd
50 (E. coli).
3. SSB (single-strand binding protein - )
DNA .
74 kd 300 (E. coli).
4.DNA (DNA ligase)
DNA. 74 kd 300
(E. coli).
5. DNA (DNA gyrase) )

( ,
, ).
400 kd 250 (E. coli).

DNA

DNA 5 3.
DNA ,
5 3 3 5.

DNA 5 3


. .

, .

Okazaki

DNA
1.000 2.000
DNA. ,
Okazaki dNTPs, rNTPs,
RNA. 24 DNA .
Coli.

H DNA .

DNA
. DNA
P ADP
. DNA
SSB
DNA.
.

. -

3 5 Okazaki.

RNA .

RNA DNA. H RNA


5 3. RNA
, DNA pol. III dNTPs 3 RNA
Okazaki, 1000-2000 .

Okazaki DNA pol. III

DNA pol. I. 5 3
RNA

Okazaki ,
DNA 3
Okazaki.

DNA

Okazaki.
DNA.
ATP ADP Pi.

DNA

3 5.

. ,
RNA, DNA pol. III
.
Okazaki

Okazaki.

DNA

DNA pol. III


.
.

DNA pol. III,



.

DNA pol. III

1000 . , ,
RNA
Okazaki.



.
:
1.- DNA
.

DNA.
,
DNA.
2.- DNA . H DNA pol.
.
RNA
. H DNA pol. .
, 3- 5
.
3.-
DNA . DNA
pol., 3-5
(de novo)
RNA 5 DNA
.
DNA.

(telomeres) - (GGGGTT Tetrahymena AGGGTT )


.

in vitro

(telomerase),
3 . GGTT, dNTPs
- GGGGTT
Tetrahymena.

GGTTAGGGTT .

Elizabeth Blackburn RNA

G-rich ( G)
.
(template-bearing reverse transcriptase).

3- G-rich DNA.
o RNA ,
- 3- ,
. 3-
.

3- . 3-

.

DNA pol.

. , DNA
3 5 , ,
.