?20? DNA ??? ????

1
?? ???? ?? ??? ?? (Understanding and
Manipulating Genomes)

• ??? DNA(recombinant DNA)
• ????(genetic engineering)
• ????(biotechnology)
• ?? 20.1DNA microarray

2
?? 20.1 2,400?? ?? ???? ?? ??? ???? DNA
microarray
3
?? 20.1 DNA ???? ?? ???? DNA ??? ??? ???? ??? ?
??? ???

• ??? ???(gene cloning)

DNA ???? ?? ????(DNA Cloning and Its
Applications A Preview)

• ?? 20.2 ?????? ??? ???? ??
• ??(clone)

??? DNA ??? ?? ????? ?? (Using Restriction
Enzymes to Make Recombinant DNA)

• ????(restriction enzyme), ??????(restriction
  site)
• ?? 20.3 ??? DNA ??? ??
• ??????(restriction fragment), ????(sticky end),
  DNA ????(ligase)

4
?? 20.2 ???? ?????? ??? ??? ???? ??, ??? ????
??? ??
5
?? 20.3 ??? DNA? ??? ?? ????? DNA ????? ??
6
?? ?????? ??? ???? ???? ???(Cloning a Eukaryotic
Gene in a Bacterial Plasmid)

• ??? ??(cloning vector)

?? ??? ??(Producing Clones of Cells)

• ?? 20.4 ?? ???? ?????? ???

?? ???? ?? ??? ??(Identifying Clones Carrying a
Gene of Interest)

• ?? ???(nucleic acid hybridization)
• ?? ??(nucleic acid probe)
• ?? 20.5 ?? ?? ???
• ??(denaturation)

7
?? 20.4 ???? ?????? ?? ??? ?????
8
?? 384?
(??? ?? ??)
(??)
9
?? 20.5 ?? ?? ???
10
DNA ???? ???? ??? ????(Stored Cloned Genes in DNA
Libraries)

• ??? ???(genomic library, ?? 20.4)
• ??? DNA(complementary DNA, cDNA)
• cDNA ???(cDNA library)

???? ???? ???? ??(Cloning and Expressing
Eukaryotic Genes)
????? ?? ??(Bacterial Expression Systems)

• ?? ??(expression vector)

????? ??? ?? ??(Eukaryotic Cloning and Expression
Systems)

• ?? ????? (yeast artificial chromosome, YAC)
• ?????(electroporation)

??? ???? DNA ?? ????????Amplifying DNA in
Vitro The Polymerase Chain Reaction(PCR)

• ???????? (polymerase chain reaction)
• ?? 20.7 PCR

11
?? 20.6 ??? ???
12
?? 20.7 ????????(PCR)
13
?? 20.2 ?????? ???? ?? ??? ??? ??? DNA? ??? ????
???
? ?????? ?? ???(Gel Electrophoresis and Southern
Blotting)

• ? ?????(gel electrophoresis)
• ?? 20.8 ? ?????
• ?? 20.9 ?????? ??? ??
• ?? ???(Southern blotting)
• ?? 20.10 DNA ??? ?? ???

??? ????? ?????? ??(Restriction Fragment Length
Differences as Genetic Markers)

• ?????? ?? ???(restriction fragment length
  polymorphisms, RFLPs)

14
?? 20.8 ? ?????
15
?? 20.9 ?? ???? ?? ?????? ????? ?????? ??
? ???? ?? ?????? ???? ??
16
?? 20.10 DNA ??? ?? ???
17
?? 20.3 ?? ???? DNA ???? ???? ? ??

• 1990? ???????(Human Genome Project) 2003? ??

???(??) ?? ???? ??? ??Genetic(Linkage) Mapping
Relative Ordering of Markers

• ?? ??(linkage map)(?? 20.11, ???)

??? ?? DNA ???????? ??(Physical Mapping
Ordering DNA Fragments)

• ??? ??(physical map)(?? 20.11, ???)
• ???? ?????(bacterial artificial chromosome, BAC)

DNA ??????(DNA Sequencing)

• ?? 20.11, ???
• ?? 20.12 DNA ???? ???
• ?? 20.13 ??? ???

18
?? 20.11 ??? ?? ??? ?? 3?? ???
19
?? 20.12 DNA ??????? ?? ???? ?? ???
20
?? 20.13 ???? ??? ?? ??? ???
21
?? 20.4 ??? ???? ??? ???? ??? ??? ????

• ???? (genomics)

DNA ????? ??? ??? ??? ????(Identifying
Protein-Coding Genes in DNA Sequences)

• ? 20.1 ?? ?? ?? ??? ??

???? ?? ????(Determining Gene Function)

• ???? ???? ?? ??(in vitro mutagenesis)
• RNA ?? (RNA interference, RNAi)

22
(No Transcript)
23
???? ?? ????? ??? ?? ??(Studying Expression of
interacting Groups of Genes)

• DNA ??????? ???(DNA microarray assays) (?? 20.14)

?? ?? ??? ??? ????(Comparing Genomes of Different
Species)
????? ??(Future Directions in Genomics)

• ?????(proteomics)
• ?? ?????? ???(single nucleotide polymorphisms,
  SNPs, snips?? ??)

24
?? 20.14 ??? ???? ?? DNA ??????? ???
25
?? 20.5 DNA ??? ???? ??? ??? ?? ?? ?? ??? ???
??? ??(Medical Applications)
??? ??(Diagnosis of Diseases)

• ?? 20.15 ?? ?? ???? ??? RFLP ????

??? ??? ??(Human Gene Therapy)

• ??? ??(gene therapy, ??? ?? ???? ??)
• ?? 20.16 ????? ??? ??? ??

26
?? 20.15 ??? ???? ?????? ??????? RFLP
27
?? 20.16 ??? ???? ??? ??? ??? ??
28
??? ??(Pharmaceutical Products)
???? ??(Forensic Evidence)

• ??? ??(DNA fingerprint) (?? 20.17)

????(Environmental Cleanup)
29
?? 20.17 ?? ?? ??? ??? ??? ???
30
????? ??(Agricultural Applications)
??? ? ?? ??(Animal Husbandary and Pharm Animals)

• ????(transgenic) ??
• ?? 20.18 ??? ??? ?? ?? ??? ??

??? ???? (Genetic Engineering in Plants)

• Ti ?????(Ti plasmid)
• ?? 20.19 ???? ?? ???

DNA ????? ?? ??? ??? ?? ??(Safety and Ethical
Questions Raised by DNA Technology)

• ??? ?? ??(genetically modified organisms, GMO)

31
?? 20.18 ?? ??
32
?? 20.19 ???? ?? ??? ?? Ti ?????? ???? ?? ??