??? ??????

1
??? ??????

• 2-1 ?????
• 2-2 ??????????
• 2-3 DNA????????
• 2-4 DNA??
• 2-5 ???
• 2-6 ????
• 2-7 DNA????
• 2-8 ?????????
• 2-9 DNA????

2

• 2-1 ?????????
• ????????????????????????????????????
• ???????????????,???????????
• ?????????????????,??????
• ???????,????????
• ????????
• ????????????,??????
• ???????
• ???????????????? ???????????????

3

• ????????????,??CO2 ?H2O???????
• ????????????,????????
• ????????????,?????
• ????????????
• ?????????
• ?????????,??(???) ????(???)

4
(No Transcript)
5
2-2 ??????????

• ???(chromosome)
• ?????DNA???????,??????(histone)
• ?????????????
• DNA??????????-??
• ???(genome)??????????????DNA

6
??????
7

• ????DNA???
• 1928 ???? (Griffith)?????????
• ??????????????
• 1942 ???(Avery)??????????????
• ????????????DNA
• 1952 ??-?? (Hershey-Chase)?????????????(S35)?DNA(P
  32),?????????
• ??????DNA????????
• 1953 ??????(Watson?Crick)??DNA?????

8
????http//fig.cox.miami.edu/cmallery/150/gene/s
f11x1b.jpg
9
????http//fig.cox.miami.edu/cmallery/150/gene/s
f11x1b.jpg
10
www.accessexcellence.org
11
www.accessexcellence.org
12

• ??????
• DNA?RNA????
• mRNA??????????RNA (snRNA)?snRNA??????(snRNPs)?SR??
  ???,???????????(splicesome)
• ???????????????
• 1970 Howard Temin?David Baltimore ?? (1975 Nobel)
• ??RNA?????????????????(RNA)????DNA

13
2-3 DNA???????
James D. Watson (left) and Francis Crick propose
a double- helical structure for DNA. (1953)
1962 ??????
(?? dna50.mpg)
Francis Crick
James D. Watson
Frederick Wilkins
14
(No Transcript)
15
??DNA????????
??????(Brenda Maddox)?????????2004?12?????
????
???.????(Rosalind Franklin, 19201958) ?????????
16
DNA X-????
17
(No Transcript)
18
DNA???
??????(nucleic acid) ??DNA(??????)?RNA(????) ???
??????(nucleotide) ?????(base)????(pentose
sugar)???(phosphate)
(????,2003)
(?? dna.mpg)
19
(No Transcript)
20
?????
Adenine
Guanine
              
Uracil
Cytosine
(????,2003)
Thymine
21

• DNA????

????http//academic.brooklyn.cuny.edu/biology
22
DNA????
????
????
????
????
????http//academic.brooklyn.cuny.edu/biology
23
2-4 DNA ??
(????,2003)
24

• DNA?????

????
???
???
????
???
???
????http//www.eimbl.org/img_file/club_file/73/dn
a_rep/index/ReplicationFork.gif
25
?????????

• ????
• ???????
• ???????
• ???????
• ????????????
• ??mRNA??????
• ???????????RNA

(????,2003)
26
?????
????http//www.nyu.edu/classes/ytchang/book/n002/
proteinsyn.gif
27

• ????

• ?????? 4 x 4 x 4 64 ???
• ????20?
• ?????????????
• ????????????
• ???????????,??????????
• ????????????

28
???
????http//campus.queens.edu/faculty/jannr/Geneti
cs/images/codon.jpg
29
2-5 ???????
????????, 2004
30
Alpha ??
31
Beta ??
32
??????????
PrPC (??) PrPSC
(???)
(Prusiner, 1999)
33
2-6 ????
??????????????
???
???
????
???? ???
(Klein et al.2003)
34
????? II
????? ??? ??????
???? ???
???
(Klein et al.2003)
35
?????????
???
???
???
? ? ? ? ?
? ? ?
??
????
??
?????
?????
(Klein et al.2003)
36
2-7 DNA ????(??????)

• ??DNA????????

???(restriction enzyme) ???(ligase) ??(vector)
? ??(transformation)???? DNA??-???? (DNA
cloning- gene cloning)
37
???

• ??DNA?????????
• ??DNA4-8????
• ??????,????

????http//www.uic.edu/classes/phar/phar331/lectu
re5/image17.gif
38
????????
????http//users.rcn.com/
39
??

• ????????????
• ?????????
• ?????(plasmid)????(bacteriophage)
• ????
• ??????
• ?????????????,???????
• ????
• ????????????????
• ?????-???????

40
??DNA?????

• 1. ???? ????-????DNA
• 
  ????-??mRNA???cDNA
• (??DNA
  complementary DNA)
• 2. ??????
• 3. ????????????
• ??????
• ?????
• ????
• ????
• 4.???????????

41
??????
(Glick and Pasternak, 2003)
42
?????
(Glick and Pasternak, 2003)
43
????
???? http//www.ufsm.br/blg220/hide/microinjectio
n.jpg http//transgenese.crchul.ulaval.ca/images/E
S20injection.jpg
44
???? I
????http//agham.asti.dost.gov.ph/1998/4th/extras
/dna4.gif
45
???? II
????http//science.marshall.edu/harrison/research
.html
46
???????????

• ??????????
• ???????
• ?????????
• lacZ?????

47
??DNA???????

• LacZ??????????,??X-gal??????
• LacZ?????DNA??,????????,????X-gal,?????

48
2-7 ?????????
???????DNA ????
mRNA ? cDNA ? ???(plasmid clones) ? ????(genomic
library) ? ??? ????(molecular probe)
49
2-8 DNA????

• ???????(polymerase chain reaction)
• ???????????DNA???

(?? PCR.mpg)
50
PCR??

• 1970 Khorana???????
• DNA?????
• ????????
• 1983 Kary Mullis??????????
• ??1993???????

51
??PCR????????

• ???DNA???(Thermostable DNA polymerase)
• Taq- Thermus aquaticus
• ??DNA??????
• ?????(proofreading)
• Pfu- Pyrococcus furiosis
• ??(primers)
• ????????? (18 32 ??)
• ????????(Deoxynucleoside Triphosphates,??dNTPs)
• ??DNA (template DNA)
• ???????

52
PCR???????

• ??(Denaturation)
• ??(95?)???DNA????
• ????(primer annealing)
• ?????????????????DNA????? (??40 72?)
• ??(extension)
• DNA?????????DNA (??????72?)
• ?????????,????25 30???

53

• PCR???????

(????, 2003)
54

• PCR???????

(????, 2003)
55

• PCR???????

(????, 2003)
56
PCR???????

• ?????????????????
• ?????????? ??DNA(???????PCR??)
• ?????
• ?????DNA??,???????5300?????????????
• (?? iceman.mpg)
• ?????????????
• ?????????
• ??????
• ??????????????

57
(No Transcript)
58
?????????

• ??????????(RFLPs)
• (Restriction Fragment Length Polymorphisms)
• ?????DNA,?DNA????, ???????
• ???????VNTR variable number of tandem
  repeat????DNA??????,???????????????????
• STRPs , simple tandem repeat polymorphisms???????
  ??
• SSLPs short sequence length polymorphisms
  ????????

59
RFLP???
(Glick and Pasternak, 2003)
60
RFLP???
????http//users.rcn.com/jkimball.ma.ultranet/Bio
logyPages/R/RFLPs_8.gif
61
?????Southern blotting

• ????DNA

????http//fig.cox.miami.edu/cmallery/150/gene/c
7.20.10.blotting.jpg
62
?????Northern blotting

• ????RNA

(??, 2006)
63
?????Western blotting

• ?????
• ???????
• ?SDS-PAGE(?????)??????????????????????

SDS-PAGE
????http//www.evidenttech.com
64
??????(in situ hybridization)

• ??????????
• ????????????

????http//www.jnanobiotechnology.com/content/fig
ures/1477-3155-4-5-1.jpg
65
DNA???

• Maxam Gilbert Method1977

66
Maxam Gilbert Method II
????http//www.cbs.dtu.dk/staff/dave/roanoke/gene
tics15.htm
67
Maxam Glibert Method III
68
Sanger's Dideoxy Method I
????http//www.cbs.dtu.dk/staff/dave/roanoke/gene
tics15.htm
69
Sanger's Dideoxy Method II
70
Sanger's Dideoxy Method III
71
Fluorescence dideoxy Method I
????http//www.cbs.dtu.dk/staff/dave/roanoke/gene
tics15.htm
72
Fluorescence dideoxy Method II