nucleic acid

1
??? ????
nucleic acid
2
????

1. ????????????????DNA????????????????????????
2. ?????????
3. DNA??????????RNA???????
4. DNA?????

3
????

• ??? ?????????
• ??? ???????
• ??? ???????
• ??? ??????

4
?????
DNA(??????)???? RNA(????)
mRNA(??RNA) tRNA(??RNA) rRNA(???RNA)
??
5
??? ?????????

• ????????????
• ??????C?H?O?N?P
• P????910

6
????????
????????
7
(?)??
RNA DNA
8
????
??? (adenine, A) ??? (guanine, G) ???
(cytosine, C) ????(thymine,T) ??? (uracil,U)

??(purine)
??(pyrimidine)
9
???(A) (6-????)
???(G) (2-??-6-???)
??
10
??
????(T) (5-?????)
???(U) (2,4-????)
???(C) (2-? ,4-????)
11
(?)? ?
12
RNA?DNA??
??
??
A G C U
D-??
??
A G C T
D-2-????
??
13
????????

• (?)???????(????)

14
(?)?????????(?? ??)
?? ?? ??
??????????5?????
15
?????????
16
(?)?????????
17
cAMP cGMP
18
??? ?????
19
??3,5-????????????
(?)???????

• ??????????3?, 5? -?????
• ?????????? 5??????
• 3?????
• ???? 5? ? 3?

20
???????
5??????
3?, 5? -?????
3?????
21
(?)???????
G C A T T
5
3
OH
P
P
P
P
P
5 p G pC pA pT pT-OH 3
5 G C A T T 3
G C A T T
22
?? DNA?????

• (?)DNA?????
• ?dAMP?dGMP?dCMP?dTMP?????????3,5-????
  ??????????????????
• ?DNA???????????????

23
(?)DNA?????

• 1 ??
• 1)?DNA???????Chargaff ??

• ?????DNA??????
• ???????????DNA???
• ????
• AT,GC,AGTC
• 2)Wilkins?Franklin???????DNA???????X???????
• 3)??????,?????????????????
• ?Pauling ?Corey??A?T??2????C?G??3????

24
DNA ?????
25
2?DNA??????????
1953?,Watson?Crick?????????
26
(No Transcript)
27
DNA???????

• ????,????
• ???????,??????????
• ??2nm,??3.4nm,10???/?
• AT,GC
• ???????
• ???????
• ?????(??)
• ??

28
(?)DNA?????

• DNA???????????????????????????????

29

• ???????
• ????DNA?????????

30
DNA???
DNA?????
????????????????3,5????????????????????
??????????????????DNA??????
??????????,DNA???????????????????????????
?????DNA???????????????(?)?
31
????DNA????????

• ???????????DNA??,????????????????????RNA?????????D
  NA?????????
• ??(gene)??????????????RNA????????
• ???????????????????,
• ?????

32
??RNA????????

• ?????????3,5-??????????????????RNA????
• ??mRNA?tRNA?rRNA

33
(?)tRNA

• ????????????????
• 1?tRNA?????
• ????
• ??????,3/???CCA-OH??
• (???????)

34
2.tRNA?????
H
????
??? uracil
(U)
35

• tRNA??????
• 4??(?????)?4??
• ??????3/???-CCA-OH,??AA
• D?
• ????(?mRNA????????)
• T ? C?
• ???(tRNA?????)

36
3?tRNA?????(?L?)
?????
??????
37
(?)mRNA
????mRNA????????,???????????????????,???????????hn
RNA? ????mRNA??5??????3??poly A?

• 1???

5 ???? ??? 3
??A?
3
5
m7Gppp AUG GUG UAA AAA An
38
mRNA?5?????m7GpppN
39

• ???????
• ??mRNA???
• ??????????????????
• Poly A?????mRNA????????
• ? mRNA??????

40

• 2?????????????
• mRNA?????????????1???????

41
(?)rRNA
90???,Noller??23SrRNA????????, ???????rRNA????

• ??
• ????????????,??????????
• ??
• ??????????????

42
31 proteins 23S RNA 5S RNA
50S
70S
??? ???
??
21 proteins 16S RNA
30S
49 proteins 28S RNA 5.8S RNA
60S
??? ???
80S
??
33 proteins 18S RNA
40S
43
(?)?????RNA
snRNA????????,???RNA???0.11,?????RNP(??????)????
?,?hnRNA?rRNA????????????????????????????????????

• ???RNA(snRNA)
• ???RNA(scRNA)
• ??(ribzyme)???????RNA
• ??RNA(asRNA)????RNA(??mRNA)???????RNA?DNA??,?????
  ??????mRNA??,??????????.?

44
??? ???????
????????? ????? 260nm
45
?A260/A280??????????
?DNA,A260/A280 gt1.8 lt2.0 ?RNA,A260/A280
gt2.0? ????????????,?A260/A280??????? lt2.0 ,
gt1.8,DNA??gt2 RNA??
? A1?, DNA50ug/ml,
RNA???DNA40ug/ml, ????20ug/ml?
????????
46
???????
1.???????????????(??????????????),??????????????,
???????????????? 2.?????????,????????????????????
???,??????????????????
47
????? ?????
48

• ?????????(???)???260nm?????( A260 )??
• ????????????260nm?????( A260 )??

49
DNA??
50DNA????
??????? Tm
(GC) (Tm 69.3) 2.44
50
?? ??????
????????????????,??????????????????????,?????????
??
??
51
??????????????,???????
??
??
?
?
?
?
?
?
?
?
?
?
?
?
52

???? p510
???????DNA?RNA????????,??????????????
Southern??????????DNA???????????????,??????
Northern??????????RNA????????????????
?????????????????????????,???????????
53
??????????? p518
?????Sanger????Maxam-Gilbert????
?????????? DNA????????? ????????DNA? ???4?????DNA
? ???,???????? ddNTP,??????? ??DNA?,????DNA ????,?
??ddNTP ??????????? ????????DNA???
54
???,??
55
(No Transcript)
56

• hnRNA (heterogeneous nuclear RNA)---????RNA
• snRNA(small nuclear RNA)???RNA
• scRNA( small cytoplasmic RNA)???RNA
• asRNA(antisense RNA)??RNA
• cccDNA(covalently closed circular DNA)????DNA
• ocDNA(open circular DNA)??DNA
• linear DNA??DNA
• dsDNA(double-strand DNA)??DNA
• ssDNA(sigle-strand DNA)??DNA

57
????

• ?????????????,?????????
• ?????U ????(??) T???????????O
  U????????U??????O??-NH2
• 
  C??????????O,???O??-NH2
• 
  G???????,????(??)?????O,G??O??-NH2???? A

58
?????? p514-516

• ?)?????
• ?)??? 9.5
• ?)?????? ???
• ????????????????
• DNA?????????????
• ????EB????

59
???? p502

• ??????????????,????????????(???DNA??????)
• ???RNA?????(??,0.1mol/LNaOH??RNA????,??2-?3-???
  ?????)DNA??
• ??? ?????????????  
• RNA??? RNase
• DNA??? DNase
• ???????????????
• ????? ???????????????????????
• ???????????? ??????????
• ????

60
Southern Blotting

• ???DNA -- ? DNA??????
• ? DNA?? --? ??????? ----
• ????? --? ??????????
• ? ?????????DNA???RNA
• ?? --? ????????

61
??????
62
?????????????????????????????,??3??????????? ????
?,?????(Escherichia coli)?Eco??,????? (Haemophilus
influenzae)?Hin??????,???????, ???????,?????????
?????,???????? ?????????????????????????d? (Haemo
philus influenzae d)??????3????, ??????Hind??Hind?
?Hind??

• ???????????
• ?????DNA????????(palindrome)
• ??DNA????5-???3-?????
• ????????5-?3-????????????????DNA??????,???????

63
???????
ß-D-2-??
ß-D-2-????
??????(???)
?
?? H
??
????
FeCl3
?
?-??-?-???
???
???? H
????
64
OH
??
???(?)
65
?????!