??????:???????????DNA??,???????????????DNA??????; - PowerPoint PPT Presentation

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Title:

??????:???????????DNA??,???????????????DNA??????;

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1 (lac operon) ... Lac O Ara O. . . ... – PowerPoint PPT presentation

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Title: ??????:???????????DNA??,???????????????DNA??????;


1
??? ???????????
2
  • ?????????????????DNA??,???????????????DNA??????
  • ????????????????(??????)?????????,
  • ?????????????????(??????)???????(???DNA?)?????????
    ????

3
  • 1.1 ??????????????????????????????????
  • ?????????????????
  • ??????????????,???????????

4
1.2 ???????
  • ?????(constitutive expression)
  • ?????(adaptive expression)

5
  • 1.2.1 ?????

???????????????????
  • ?????????????????????,?????????(housekeeping
    gene)?
  • ??????????????????
  • ????????????????????????????????????

6
  • 1.2.2 ?????
  • ????????????????????????
  • ??(induction)??????????????????,?????????????(ind
    ucible gene)
  • ??(repression)?????????????????????????????????(r
    epressible gene)?

7
1.3 ??????? ???????
  • ??????????-?????????????,???????????????????
  • ???????????????????,??????????????????
  • ??????????-????????????????,?????????????
  • ??????????????????????

8
1.4 ????????????
  • ????????????(?????)
  • ?????????(?? )
  • ??????????????,????????????,?????????????????,????
    ??
  • ??????????????????,????????????????????

9
  • 1.5 ????????????
  • ? ????????
  • ? ?????????
  • mRNA??????????
  • ???????? ?

10
  • ??? ??
  • ??? ???
  • ??? ????????
  • ??? ???????

11
2 ????
  • 2.1 ?????????
  • 2.2 ????
  • 2.3 ???????
  • 2.4 ?????????????
  • 2.5 ????

12
2 ????
  • 2.1 ?????????
  • ????(structural gene)??????RNA??????
  • ???????????????,?????????
  • ????(regulator gene)???????????RNA??????????
  • ??????DNA???????????????

13
(No Transcript)
14
  • 2.2 ????
  • ????(operator gene,O)????????????DNA?????????
    ????????,?????????????


15
???????????????????,????????????,?????????????
16
???????????????????,????????????,??????????????
17
2.3 ???????
  • ??? (promoter, P) ??RNA??????????????????DNA???
  • ???(terminator, T) ??RNA??????????DNA???

18
2 ????
  • 2.1 ?????????
  • 2.2 ????
  • 2.3 ???????
  • 2.4 ?????????????
  • 2.5 ????

19
  • 2.4 ?????????????
  • ??????(cis-acting element) ???????????DNA??,?????
    ??????????DNA??????
  • ????????,????????????,
  • ???????????????

20
  • ??????(trans-acting factor) ??????????(??????)???
    ??????,
  • ?????????????????????DNA????
  • ????????????

21
2.5 ????
  • ???????????,RNA????????????
  • ?????????????????????????,RNA??????????
  • ???????????????????????????

22
TBP
Pol III ? ? ?
TFIIIB
TFIIIC TBP
RNA pol
III

Pol I ? ? ?
RNA
pol I SL1
UBF1
TBP
Pol II ? ? ?

?????
TATA

TFIID
TAFs
RNA pol II
23
  • ?????????????????(??????)???????(???DNA?)?????????
    ????

24
  • ??? ??
  • ??? ???
  • ??? ????????
  • ??? ???????

25
??? ???
  • 1 ?????(lac operon)???
  • 2 ??????????
  • 2.1 ?????????
  • 2.2 ?????????
  • 2.3 ??????????????
  • 3 ??????
  • 4 ???????????

26
  • 1 ?????(lac operon)???
  • ??1940?Monod??????????????????,?????????,??????,
    ???????????
  • 1961?,???Jacob Monod??????????
  • ?1965????????????

27
????
??
??
?(?)?
?(?)?
????
ß-?????
???
??
???
28
T
  • ????????
  • ????? Z Y A
  • ??? P
  • ????O
  • ???? I
  • ??? T

29
?????????????,???????????????????????????????????
???????????
30
??? ???
  • 1 ?????(lac operon)???
  • 2 ??????????
  • 2.1 ?????????
  • 2.2 ?????????
  • 2.3 ??????????????
  • 3 ??????
  • 4 ???????????

31
1 ?????????
  • ß-?????????????????
  • ?????????????,??????????????????

32
  • 1.1 ????????-?????????
  • ???(inducer)???????-????,?????????????

33
  • 1.2 Lac??????????
  • ????????????????
  • ????ß-???????????????????????
  • ??
  • ????ß-???????????
  • ??????Lac????????
  • ?????????????????????

34
(No Transcript)
35
  • ?????
  • ???????????????????????,????????????,?IPTG(???-
    ß D-??????)?

36
??? ???
  • 1 ?????(lac operon)???
  • 2 ??????????
  • 2.1 ?????????
  • 2.2 ?????????
  • 2.3 ??????????????
  • 3 ??????
  • 4 ???????????

37
(No Transcript)
38
2 ?????????
  • ??????????
  • ???????DNA(????)?????
  • ?????DNA???????
  • ?????RNA??????

39
2.1 ?????????????
HTH
??
???
  • N?159aa,????HTH,?????DNA?????
  • 2???????6????,?????????????????
  • C????a-??

?????1
?????2
??-??
40
4???????????????
41
2.2 ???????DNA(????)?????
???11
?????????
42
  • 2.3 ?????DNA???????
  • ??????,????????DNA?,?????????,???????????
  • ?????????????????

43
  • 2.4 ?????RNA??????
  • ?????RNA pol????DNA??
  • ?????????RNA pol??? RNA pol ???????????
    1.9X107??????,2.5X109?
  • ??????????RNA pol????????????,???????,????????????
    ?,???????

44
???????????????
45
??? ???
  • 1 ?????(lac operon)???
  • 2 ??????????
  • 2.1 ?????????
  • 2.2 ?????????
  • 2.3 ??????CAP????
  • 3 ??????
  • 4 ???????????

46
2. 3 CAP????????????
  • 2.3.1 CAP (?????????,catobolite gene activatorin
    protein)
  • ?cap??,??cAMP???????CAP
  • cAMP??????,??????????????

47
(No Transcript)
48
??????????????cAMP??,????cAMP-CAP???,?????
cAMP-CAP????lac???????
49
2.3.2 CAP?????
  • (1)CAP??????????
  • a ????CAP???RNA pol a??,
  • b ???DNA,?????,????RNA Pol???

50
(2) CAP????
  • ????22bp I -70 -50
  • II -50 -40
  • CAP????, 45KD ,?cAMP??

51
2.3.3 CAP???? ??pLac?????,??????????????lac???
????,????????????,?????CAP???????,????????????????

52
(No Transcript)
53
??? ???
  • 1 ?????(lac operon)???
  • 2 ??????????
  • 2.1 ?????????
  • 2.2 ?????????
  • 2.3 ??????CAP????????
  • 3 ??????
  • 4 ???????????

54
3 ?????? 3.1 ???trp??????
55
3.2 trp??????????
  • 1) ????????

???????,??????
56
???????- trp???
  • ??????????,?LacR?1/1000
  • ????(corepressor)???????,???????????????
  • ???(inducer)???????-????,?????????????
  • ???(effector)??????????????????????,?????????????
    ?,???????????????????

57
2) ????/????
  • ??????????,?????????????????????????,?????????????
    ????,??????????????????

58
  • ???? trp L (leader sequence) ?trp mRNA
    5?trpE???????????162bp?mRNA????????????????????
  • ???(attenuator) ???????, DNA??????????????????(12
    3-150bp)?
  • ????(attenuation) ?????RNA????????????????

59
(No Transcript)
60
????
61
  • ??????????
  • ???Trp ???,????????????????,????mRNA???
  • ??????????
  • ?trp????,?????????????,????,??????trp
    ???????????????????,??????????Trp???
  • ??????,????????
  • ???? ???trp?,???
  • ???trp?,???????
  • ???? ??
  • ?????????????,?????????????

62
(No Transcript)
63
??? ???
  • 1 ?????(lac operon)???
  • 2 ??????????
  • 2.1 ?????????
  • 2.2 ?????????
  • 2.3 ??????CAP????????
  • 3 ??????
  • 4 ???????????

64
5 ???????????
  • ????????????
  • ????????????????,
  • ????????
  • ????????
  • ????????????????,
  • ????????
  • ????????

65
(No Transcript)
66
(No Transcript)
67
  • ??? ??
  • ??? ???
  • ??? ????????
  • ??? ???????

68
??? ????????
69
1 ?mRNA??????? ?????mRNA??????,?????????mRN
A?????????????????????
rpIL(L7)
70
(No Transcript)
71
2 ????????rRNA
  • ?????rRNA16S,23S?5S?
  • 7??????rRNA,rrnA-G,????????????
  • ????????16S?23S?5S RNA???tRNA?tRNA???,??????????
  • ????????????????
  • ???????16S rRNA?????300bp?,???????????P1
    110bp???????P2?

72
  • ??? ??
  • ??? ???
  • ??? ????????
  • ??? ???????

73
??? ???????
  • ??????????
  • ??????????
  • ????????
  • ?????????????
  • ???RNA????

74
1 ??????????
  • ???????????RNA????????
  • 1.1 ????????????
  • 1.2 ????????????????????
  • 1.3 mRNA??????????

75
  • 1.1 ????????????
  • ???????mRNA???????,???????????????
  • ?????????????????AUG???????,??????????

76
? ?mRNA??????????????
77
T4?????p32???????
p32????DNA??,??????? ?
????DNA,???p32??????mRNA??????????
? ?????32 ???p32????mRNA???????????
78
  • 1.2 ????????????????????
  • ??????????????
  • ???????????????RNA???????

79
?
80
  • ????????????????????????,?????????????
  • ??????????????????????????
  • ???????????????????????????

81
1 ??????????
  • ???????????RNA????????
  • 1.1 ????????????
  • 1.2 ???????????????????
  • 1.3 mRNA??????????

82
  • 1.3 mRNA??????????
  • ????????????????,????????????????????????
  • mRNA??????????,????????????mRNA???????,???????????
    ????????????

83
(No Transcript)
84
??-RNA??????????
CP ????
Rep???
CP?Rep????
85
??? ???????
  • ??????????
  • ??????????
  • ????????
  • ?????????????
  • ???RNA????

86
(No Transcript)
87
2 ??????????
  • ??????
  • TrpE-Thr-Phe-?????
  • ACU UUC UGAUG GCU
  • Met-Ala-TrpD
  • TrpB -Glu- Ile-??
  • GAA AUC UGAUG GAA
  • Met-Glu-TrpA

88
??? ???????
  • ??????????
  • ??????????
  • ????????
  • ?????????????
  • ???RNA????

89
3 ????????
  • ??????,????????????????????,???????????????,??????
    ????,?????????

90
(No Transcript)
91
  • ? ??????Ile?????????
  • ??????????,????????mRNA?????????tRNA?????????
  • ??tRNA????????????????,?????tRNA??????????????

92
??? ???????
  • ??????????
  • ??????????
  • ????????
  • ?????????????
  • ???RNA????

93
4 ?????????????-????
  • ????(strigent control)????????,????????????????,?
    ??????????????????,??????????
  • ???????, tRNA?????,???ATP?GTP????????,???????ppGpp
    ?pppGpp?????????????????,?????
  • GTPATP ? pppGppAMP?ppGpp
  • ?????????rRNA?tRNA????? ?

???(????RelA)
94

(????RelA)
( spoT ?????,??ppGpp??????)
95
  • ???????
  • pppGpp????????????,????RNA????????????????????????
  • ??????????????,
  • ???????????????,
  • ???????????????,
  • ?????????

96

(????RelA)
( spoT ?????,??ppGpp??????)
97
??? ???????
  • ??????????
  • ??????????
  • ????????
  • ?????????????
  • ???RNA????

98
  • 5 ???RNA????
  • 1984?Mizuno?Iwoue??RNA?????????,???????,?????????
    ?
  • ?????RNA (small interfering RNA, siRNA)??RNA????
  • ????????????
  • ?????RNA???????,?????,??????????????????????

99
  • ?????RNA????????????
  • RNA???????,????????????????????
  • ?????????????,?????????
  • ??RNA ?mRNA???RNA???
  • ??RNA?????????RNA -RNA????

100
  • ??RNA???????????
  • ?mRNA??????????,????????
  • ?mRNA???????,???????
  • ???????,????????

101
??-E coli ? ompF ?????????RNA??
EnvZ
EnvZ,?????????????
ompF?????, ??????????
102
(No Transcript)
103
  • ??RNA??????????,???????RNA,?????????mRNA????
  • ???????????????????,??????RNA?????????
  • ????????????RNA???RNA????????????RNA??mRNA?????,
    ????????????????????

104
???????????
  • ACC, 1-??????????, ????????
  • PG,??????
  • ???????????

105
???
  • ?????????????????????? ?????????????
  • ???????????
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