The Popular Technology in Molecular Biology: Principle and Application

1
?????

• ???????????????
• The Popular Technology in Molecular Biology
  Principle and Application

2
? ? ??????????Molecular Hybridization
Blotting Technology
3
??????????????
?????? (nucleic acid hybridization )
?DNA?????,?????DNA???????????,??DNA?RNA????,???DN
A?RNA?????????????????,?????????????????(heterodup
lex) ?
4
??
RNA
DNA
5
(?)???? (?)????
?? (probe) ????????????????????????????????????
,????NC????????,???????????????
6
????????????
(?)DNA???? (Southern blotting)
?????DNA?????????????
(?)RNA???? (Northern blotting)
??RNA????????
(?)???????? (Western blotting)
?????????????????
7
?? ???? (dot blotting) ???? (in situ
hybridization) DNA?? (DNA array) DNA????
(DNA chip)
8
?????????
9
? ?
10
???????
? ?
11
DNA??
? ?
12
? ? ? ? ? ? ? ? ?Polymerase Chain Reaction
13
????????
Template DNA
Primer 1
5?
5?
5?
Primer 2
5?
? ?
14
? ?
15
??PCR????????

• ??DNA
• ?????
• ??DNA???
• dNTPs
• Mg2

16
??PCR???????
?? 95C
?? Tm-5C
?? 72C
17
??PCR?????
(?)??????? (?)??????? (?)DNA?RNA????? (?)DNA???? (
?)??????
18
???????PCR????

• (?)???PCR??
• (?)??PCR??
• (?)??PCR??

19
??PCR????
? ?
20
? ? ? ? ? ? ? ? ?Nucleic Acid Sequence
Analysis
21
???????????
????? (Maxam-Gillbert?)
DNA????????? (sanger?)
22
???????(Maxam-Gillbert?)
????
???????????DNA??1??2??????????????,?????????,?????
??????????,???????????,????????????DNA??,?????????
?? ???,??????DNA?5??,?????????X?????DNA?????
23
??DNA????????
24
? ?
25
??DNA????
????????????????DNA????????????????????????????,??
??Sanger????,???????(??????????)???,????????????DN
A?????????????????????,?????????,?????DNA????????
26
DNA????????
? ?
27
? ? ? ?Gene Library
? ? ?
28
???? (gene library)
??????????????DNA????????

• ???DNA?? (genomic DNA library)
• cDNA?? (cDNA library)

29
???DNA??
? ?
30
???????????
?????
?????
?????
31
????????????Cloning and Identification of
Disease Relative Genes
? ? ?
32
???????????
(?)?????(functional cloning) (?)????(positional
cloning) (?)???????????(position-
independent candidate gene approaches) (?)????????
??(positional candidate gene
approaches )
33
(?)?????
?? ????????????????,????????
?? ?????????????????????????????
34
???? ? ????????????cDNA?? ?
?????????????????????,??cDNA???
?????? (functional complementation
assay) ???????????????????
35
(?)????
?? ?????????????????????,????????
????????? ? ?????(???????????) ???????????? ?
??????? ?????(??????)???????????????
36
(?)???????????

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

37
(?)??????????
??????????????,????????????????????????,??????????
38
? ? ???????????????
The Establishment and Application of
Heredity-Modified Animal Model
39
???????
????? ???????????????????????????,???????????,????
????
??????????? ?????(transgenic animal) ?????????
40
???????

• ?????
• ?????????,????????????????????????,???????,???(clo
  ne)?

41
? ?
42
????????
?????? ??????(gene targeting)??,?????????????????
43
????????????????????

• ??????
• ? ?????????
• ????
• ?????(gain-of-function mutation)
• ? ?????????

44
? ? ?          ? ? ? ? ? ?
Biological Chip Technology
45
??????(gene chip)

• DNA??(DNA chip)
• cDNA??(cDNA chip)
• ????????DNA???cDNA??????,???????????????????? ?

46
? ?
47
???????
?????(protein chip)
????????????????????????????,???????????,?????????
?,?????????????????????????
48
? ? ? ???????????
Research Technology of Interaction of Protein
49
???????????????
??????????????????????????????????????????????????
???,??DNA??????????????????????????,??????????????
?
50
??????????????
????? ??????(???????????) ???????????? ???????????
51
??????????????
? ?
52
 ????????????
(?)????????????? (?)?????????? (?)?????????? (?)??
??????????? (?)?????????