The Popular Technology in Molecular Biology: Principle and Application

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?????

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

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? ? ??????????Molecular Hybridization
Blotting Technology
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??????????????
?????? (nucleic acid hybridization )
?DNA?????,?????DNA???????????,??DNA?RNA????,???DN
A?RNA?????????????????,?????????????????(heterodup
lex) ?
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??
RNA
DNA
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(?)???? (?)????
?? (probe) ????????????????????????????????????
,????NC????????,???????????????
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????????????
(?)DNA???? (Southern blotting)
?????DNA?????????????
(?)RNA???? (Northern blotting) ??RNA????????
(?)???????? (Western blotting)
?????????????????
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?? ???? (dot blotting) ???? (in situ
hybridization) DNA?? (DNA array) DNA????
(DNA chip)
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? ?
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???????
? ?
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DNA??
? ?
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? ? ? ? ? ? ? ? ?Polymerase Chain Reaction
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????????
Template DNA
Primer 1
5?
5?
5?
Primer 2
5?
? ?
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? ?
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??PCR????????

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

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

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

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??PCR????
? ?
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? ? ? ? ? ? ? ? ?Nucleic Acid Sequence
Analysis
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???????????
????? (Maxam-Gillbert?)
DNA????????? (sanger?)
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???????(Maxam-Gillbert?)
????
???????????DNA??1??2??????????????,?????????,?????
??????????,???????????,????????????DNA??,?????????
?? ???,??????DNA?5??,?????????X?????DNA?????
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??DNA????????
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? ?
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??DNA????
????????????????DNA????????????????????????????,??
??Sanger????,???????(??????????)???,????????????DN
A?????????????????????,?????????,?????DNA????????
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DNA????????
? ?
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? ? ? ?Gene Library
? ? ?
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???? (gene library)
??????????????DNA????????

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

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???DNA??
? ?
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???????????
?????
?????
?????
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????????????Cloning and Identification of
Disease Relative Genes
? ? ?
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???????????
(?)?????(functional cloning) (?)????(positional
cloning) (?)???????????(position-
independent candidate gene approaches) (?)????????
??(positional candidate gene
approaches )
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(?)?????
?? ????????????????,????????
?? ?????????????????????????????
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???? ? ????????????cDNA?? ?
?????????????????????,??cDNA???
?????? (functional complementation
assay) ???????????????????
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(?)????
?? ?????????????????????,????????
????????? ? ?????(???????????) ???????????? ?
??????? ?????(??????)???????????????
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(?)???????????

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

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

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

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????????
?????? ??????(gene targeting)??,?????????????????
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????????????????????

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

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

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

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? ?
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???????
?????(protein chip)
????????????????????????????,???????????,?????????
?,?????????????????????????
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? ? ? ???????????
Research Technology of Interaction of Protein
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???????????????
??????????????????????????????????????????????????
???,??DNA??????????????????????????,??????????????
?
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??????????????
????? ??????(???????????) ???????????? ???????????
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??????????????
? ?
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 ????????????
(?)????????????? (?)?????????? (?)?????????? (?)??
??????????? (?)?????????