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??? DNA????????

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Title: PowerPoint Author: zyl Last modified by: yyl Created Date: 4/28/2000 11:34:25 AM Document presentation format: – PowerPoint PPT presentation

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Title: ??? DNA????????


1
??? DNA????????
2
?????????? ??? ?????????? ??? ???? ???
?????
??? ???? ??? ????(????) ??? ????????
3
??????? ? ?????(??????????????????
?????? ????????DNA???) ? ?????????(????????)
? ?????????(?????????) ???????
???????(???)???????(????
?)??????????????????(???? ?)????????
4
??????? ? DNA????????(?????) ?
???-N-??????? (???U????C?????U) ?
?????? ? ??????(?????????SOS???)
? ??????-----????DNA???
5
??? ???????????
6
1??????????????????? ?????????102103?
??????(MRS Mismatch Repair System)
DNApol (? 10-8) ??????? 10-11
7
2???????(Mismatch repair system)
(1)??
DNA polymerase Helicase SSB ????? (???)
???
MCE (mismatch correct enzyme) 3
subunits mutH, L, S
8
? DNA???????????
DNA??GATC(palindromic seq.) ?m6A???????
???2kb????GATC seq.
?????????????
9
(2) ????
a?MutH/MutS ?????? ??????GATC??
??--???GATC? G?5?
????????
DNA helicase II, SSB,
exonuclease I????? ??????
DNA polymerase III ? DNA ligase ????
?????????DNA????
10
???????????5?(????????5?) ----------------?----
------------3?(--------------------------3?)
11
3? ???-N-????? ( ung system )
????????dUTP???
??????????
12
---TAGC--- ---A CG---
G
C
U
A
U
---TAGC--- ---ATCG---
---TAGC--- ---A CG---
---TAGC--- ---ATCG---
13
??? DNA?????
14
??????????
???????
???????
15
(No Transcript)
16
??????????
1????(photo reactivation )
  • ?????????
  • 400 nm ???PR ?
  • (photo-reactivation enzyme)
  • ?????(photolyase)

----TT---- ----AA----
17
2. ???? ExisionRepair
??????
  • ?????
  • ????

???????
  • UvrA, B, C gene
  • ?????
  • (Endonucleases)
  • ?????
  • (Exonuclease)
  • DNA pol
  • Ligase

18
?
?
???
?
Pol? ???
???
?
19
3. ????(RecombinativeRepair)
??????E.coli?????
??????????
20
? Rec-A. gene ???????DNA????
? Rec??????????????
???? ? Uvr????????? ?
Rec??????????????? ???????
21
? ???????
22
??????
? ?Rec-A???????(strand transfer)??
? TT dimer????,?????????TT dimer
?????
? ???????,?????????
23
???? (?????)
  • ?????
  • ????

??????
  • RecA, DNApolymerae
  • ligase

RecA
??????????? ??????????
???????
24
4. ????(SOS?? SOS repair)
(1) ????
? 80 10
100 mut. 100
50 10
UV ??? W??(Jean Weigh)
  • ?????? DNA ?E.coli A???
  • E. coli ?SOS????U.V.?? (A B)
  • SOS ?????????????(???)

25
(2) SOS ????
SOS ??--??????DNA??
?????????,????????
SOS????,???????????????
26
SOS ????SOS??????
RecA?SOS?? ????????
RecA?LexA?????
RecA?LexA??????
27
RecA-P ????
28
?DNA????/ DNA damaged
29
  • ???????
  • 1???????????
  • ??????
  • ???-N-???????
  • 2?DNA??????
  • ?????
  • ?????
  • ????
  • ????
  • ????(SOS??)

30
?DNA???????
31
??? ?????
32
1? ??-????(restriction and modificaion)
50??? Luria(T?????) Bertani(?)
Weigle(P2???)
33
E.coli K??? B????????????? E.coli C????
??
? ????????????????????DNA?? ?
?????DNA????????? ??????-????
???????-?????????DNA???DNA
34
??DNA ???????DNA
????????DNA????????????? ??DNA(resident
DNA)????????????DNA,
????DNA,??DNA,???DNA?
35
2???-????
??????????(???)
36
??? ????
37
??(mutation)??????????DNA??
????????
????
? ?????(Chromosome mutation )
chromosome number chromosome structure
? ?????( dNt point mutation )
38
???(mutant)???????????????
????(mutantgene) ?????????
?????(wild type gene)
???? ??? Arg Arg- ???
arg arg- ??? ???
39
1??????????? ????--??????????????
?? --???????
? ???????DNA??????? -----????
?????
40
???????DNA?????
41
???????DNA?????
G(k)
C(i)
C(a)
C(i)
C(a, anti)
G(k, syn)
G(k, syn)
G(k, anti)
42
2??????????? ????(point mutation) (???)
????(multiple mutation)
???---??????????????
? ????( conversion )
43
3????????????
????(frameshift mutation)??????????

???,?????????
????
? ???????? (dNt deletion or insertion )
44
Examples of deletion mutations
45
4?????????????(???) ???????????????????
??????????????????????
(?????????) ??????????????????????
???????????
? ????????????
46
??????
UAA (Oc) ???(Ocher)
UAG(Am) ???(amber)
UGA(Opal) ???(Opal)
47
5????????? ???
---??????
---?????
????? ????? ????
(?,?????)
6????????? ???? ?????????????????????????
?? ??????????? ????????????----????
48
7??????????????????
??????? ???????
???????????????? ???????
??????????? ????? ?????????????????
??????
???????????????? ??????????????????
?????????????????
49
??? ???? (????)
50
wild type
mutant (???)
1??????????
?????????????????????? ????????????????????????
???
51
2????????????
?????? ??????????????????
????????????
?????? ?????????????????,?
??????????????????
????????????,???????
52
3??????????
a) AGC (Ser) ? ACC (Thr) ? AGC (Ser)
b) AGC (Ser) ? AGG (Arg) ? AGT (Ser)
c) AGC (Ser) ? AGG (Arg) ? GGG (Gly)
if
Ser Gly
53
4??????????
(1)??????? (Intragenic suppression)
???? ????
???????????? ?????????????
??????????????--- ?????????????????
(?????????)
54
a?????
55
(No Transcript)
56
----UGG 174-----------GGA210----
C
(C) (G)
----UGG174----------GGA210----
A
(K) (E)
????
----UGC174-----------GAA210----
57
  • ???????
  • 1????????
  • 2??????
  • ??????? ??????? ???????
  • ?????????????????????
  • ?????????????????
  • 3? ????
  • ???????????
  • ???????????????????
  • ??????????????

58
(No Transcript)
59
(2)????????
????---?????? ????---?????? ????---??????
??? tRNA????tRNA????????
60
a???????????( Nonsense suppressor )
???
UAG?UGA?UAA---?????? 50
???????tRNA???????,???????
61
b???????????( Missense suppressor )
62
c??????????
????????????(??)????????
??????(??)???????
63
??? ????????
64
1??????(Base analog)
2-???? 2-Amino purine
5-???? 5-Bromine Uracil
65
G
A
AT?GC ??
5-BrU
?????? GC?AT ??
66
??????????????
67
2? ???????????
??????? ???(nitrous acid HNO2)
??(hydroxylamine HA) ?????(ethyl
mathanesulfonate EMS) N-??-N-??-N-????
(N-mathyl-N-nitro-N-nitrosoguanidion NNG)
??---????
68
NH2OH (Hydroxylamine HA ??)
69
3??????????
??? (Acridine Orange AO)
??????
???? (Ethidium Bromide EB )
????
-ATTTTTCG - -TAAAAAGC-
-AT EB TTTTCG- -TA
AAAAGC-
X
????---????
70
4? ??????????
5? ????(mutator gene )
??????????????????????? ????,??????????????
????
71
??????
72
6??????????
73
??????????????????
74
7? ????(Mutation Hotpoint)
?????????????
DNA?????????????????,???????----
75
a) ???? (repetitive seq. )/ palindromic seq.
b) m5C
???????????????, ???????????? ????????50
c) ???????????,?????????
76
DNA ????????
Mutagen (???)
???????
DNA ??
?????
????
??
????
77
  • ????
  • ???? ???? ???? ???? ?????
  • ??DNA ????? ????
  • ???
  • 1?????????????????????????
  • 2?m5C??????????
  • 3??????????????????
  • 4?????????????????
  • 5????-N-?????????????????
  • 6????????????????????????
  • 7???RecA??,??SOS????????
  • 8?????????????????

78
????!!
79
  • 2003-2004???? 6??
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