2 1. (a Unit of Inheritace) RNA DNA 2. (Nature of gene) heredity heredity & variation variation

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1 Structure & Organization of Genes The Key Laboratory of Molecular Medicine Ministry of Education Weihh

2 2 1. (a Unit of Inheritace) RNA DNA 2. (Nature of gene) heredity heredity & variation variation

3 3 Concept of the gene

4 (two basic questions) 4 (central dogma) Structure Function

5 5 v 1 DNA v 2 & 1 2 v 3

6 6 DNA Composition and

7 DNA 3. DNA 4. DNA 5. DNA 6. DNA

8 DNA RNA 8 ( ) A G C T =

9 9 二 DNA的一级结构核苷酸的排列顺序一级结构 4种核苷酸的连接及其排列顺序 C 磷酸碱基脱氧核糖主链磷酸脱氧核糖侧链碱基磷酸脱氧核糖 T A G

10 DNA 10 A, G, C, T AGGACCTTGAAGGTTCCAAATTGG 01 DNA DNA

11 DNA 11

12 12

13 (base match) 13 A=T, C G

14 DNA 14

15 B-DNA 15 v 0.34nm 10 bp 3.4nm 34Å v DNA v C G CG C2

16 A form B form Z form 16 DNA RNA 0.256nm 0.28nm 0.34nm 1 11bp 3.4nm 1 10bp CG CG C3 CG CG G C2 C3 G 0.37nm; 4.46nm 1 12bp C C2 G

17 17 34Å

18 z-dna 18

19 DNA 19

20 20 DNA

21 21 DNA 174 DNA 5 -(T X G Y ) n (A X C Y ) n -3

22 22

23 DNA (T X G Y ) n (A X C Y ) n DNA telomerase 1. Azzalin, CM; Lingner, J, Telomeric Repeat Containing RNA and RNA Surveillance Factors at Mammalian Chromosome Ends. Science, Vol no. 5851, pp Azzalin, CM; Lingner, J, Telomere wedding ends in divorce. Science, Vol no.5667, pp

24 6 DNA 24 v denaturation denaturation v renaturation renaturation annealing annealing DNA

25 Polymerase Chain Reaction, PCR 25

26 26 Molecular hybridization

27 DNA 27 v v v Tm 50%DNA

28 28

29 29 GC AT

30 Tm 30 DNA Tm Tm v 1 DNA v 2 DNA G+C G+C v 3

31 DNA DNA Tm DNA 3. DNA M Na + 5.

32 32 Genome & Genome Organization

33 33 v 1 (gene) v 2 (genome) v 3 (genome organization)

34 34

36 X174 Open Reading Fram, ORF A B

37 1.2 37

38 mrna X174 PD

39 39 λ

40 1.5 40

41 41 λ

42 42 v 1 v 2 Operon Promoter mrna 600 v 3 v 4 v 5 trna rrna RNA rrna trna

43 lac Operon 43 trp Operon

44 44

45 trna rrna 45

46 46 v 1 v 2 v 3 v 4 v 5

47 47

48 48 v 1 30% <3% v 2 70% 3%

49 49 v kb v 10 v 20

50 50 <3% 3~

51 1 51

52 2 52, interrupted gene

53 53 DNA mrna 8 mrna 7 7

54 (intron) (exon) 54 v (intron) v (exon):

55 55

56 56

57 57 GT/AG v : 5 GT,3,3 AG AG 5 3 GT AG

58 3 58

59 59 simple eukaryotic transcription units mrna complex eukaryotic transcription units mrna,

60 1 60

61 2 mrna 61 ( RNA) mrna. a b T

62 62 v 1 v 2 Splicing Form v 3

63 63 v v

64 4 gene family 64 v 25% e.g. v 2. v

65 4 65 v 1. v 2. v 3. v 4.

66 : rrna trna

67 4.1.1 rrna 67 v 28S 18S 5.8SrRNA v 5SrRNA 1

68 68 1 5SrRNA 2000

69 4.1.2 trna 69 v v v trna

70 DNA 27% 66% RNA 6% ( )

71 71

73 v GH CS PRL v

74 74 v v v v v

76 76 ε δ HS

77 77

78 v src 250 AA v DEAD 8 AA Asp-Glu-Ala-Asp

79 4.4

80 Fab Fc Fc

81 79 v 10 9 v bp

82 v Ig combinatorial joining v V C D J (junctional flexibility) v (somatic hypermutation)

83 4.4.3 Ig 06 v v IgG IgA IgM IgD IgE Ig 2

85 08 v v

86 09 100~

88 11 v v

89 K 12

90 13

91 14 V H ND H NJ H V

92 class switch 15

93 16

94 17 B 10 5 V-D-J B

95 5 pseudogene 18 duplication v v v v ( )

96 19

97 1 DNA 20 v GC v GC

98 21 v 1 (satellite)dna 5~100bp,, 100Mbp. v 2 (minisatellite)dna 6~70 bp,, 0.5~30kb,, variable number tandem repeat, VNTR v 3 (microsatellite)dna 1~8bp, (simple tandem repeat, STR),

99 22 2 (minisatellite)dna: v DNA GC 10~15bp v DNA Southern blot ~1000 DNA v DNA

100 23 3 (microsatellite)dna : v 10~50kb 1 DNA STR ( ),.

101 2 24 DNA transposon DNA - cut-and-paste retrotransposon RNA DNA LTR LINE SINE

102 25 v 1 LTR (long terminal repeat) v 2 LINE (long interspersed nuclear elements ) mrna 6.1kb 3500 v 3 SINE (short interspersed nuclear elements ) LINE Alu

103 Alu 26 DNA 130bp 170bp 30-50

104 27

105 28 DNA polymorphism

106 29 v DNA 99.9% 0.1% 0.1% DNA v DNA DNA DNA 1 DNA

107 3.1 DNA 30 v RFLP v (TR) v (SNP)

108 3.1.1 Restriction Fragment Length Polymorphism, RFLP) 31 v RFLP DNA v DNA 10 v

109 32

110 v ( ) ( VNTR) v ( ) (STR)

111 ( ) VNTR) 34 v Variable Number of DNA 15~65bp 1 2 3

112 35 v DNA v

113 (STR) 36 v 1~8bp 2 3bp v DNA 2~4bp v

114 37 10 CA 12 CA 9 CA 10 CA 9 CA

115 3.1.3 Single Nucleotide Polymorphism,SNP) 38 SNP v SNP v ACTGGCATTAACCCATT AGTGGCTTTAACCCATT v 1000bp SNP

116 SNP 39 v / v v SNP SNP

117 %

118 SNP 41 v SNP DNA v 300~1000 SNP v HapMap

119 HapMap 42 v. v (Haplotype) SNP. v SNP (Haplotype Map, HapMap)

120 HapMap v v v v,

121 3.2 DNA 43 v 1 v 2 I II

122 44

123 45 X X 1/1200 1/ % 20%

124 X FMR1 46 FMR1(Fragile X Mental Retardation-1) X FMR1 5 - (CGG)n v v v 1 n 7~ n 60~230 FMR1 3 n 230 FMR1

125 e.g

126

127 v RFLP Huntington s disease HD v v

128 50 v IT15 4P6.3 HD v CAG 6 35 > HD 17 v HD v HD

129 RFLP 51 v v RFLP G8 Southern blot 4 4 v G8 17.6kb 4 v G8 Huntington s disease RFLP G8 HD v 4

130 GAA Glu GUA Val

131 DNA DNA typing 53 v DNA DNA v RFLP STR VNTR SNP v 1. DNA 2. PCR STR/ VNTR 3. SNP

132 DNA 54 VNTR DNA

133 个体识别鉴定罪犯 55

134 PCR-STR 56 STR STR STR bp DNA PCR DNA

135 57 v proto-oncogene oncogene akt ets H-ras, hst-1 v anti-oncogene Rb p53 APC v

136 (The Cancer Genome Atlas, TCGA) v DNA,, v

137 / 10 / 100 v

138 Pharmacogenomics) 59 v

139 60 v SNP SNP v

140 61 v Herceptin HER-2 25% 30% Herceptin

141

142 63 Part III Human HGP

143 64 DNA

144

145

146 1. genetic map 66 1 (cm,centimorgan)

147 67 v 1 ABO HLA 70 RFLP v minisatellite core VNTR 1989 microsatellite marker STR v MIT Lander ES SNP single nucleotide polymorphysm

148 2. map 68 DNA STS bp

149 69 v STS DNA contig DNA ,000 STS DNA

150 70 v DNA contig v v (sequence-tagged site, STS) DNA DNA v CpG Alu isochore v (cytogenetic map) v cdna/est

151 3. sequence map 71 DNA DNA DNA

152 72 1 (contig] 2 3 [sequencing]

153 4. gene map 73 cdna / ESTs expressed sequence tags 20

154 74 v v v

155 75

156 v 2.91Gbp v 300 v % v 1/1250bp

157 v v v v v " " " " Y 200 SNP 30

158 76 (post-genome eva) v post-genome project

159 1 DNA 77 v gene targeting knock out knock in v gene silencing RNA mrna RNA RNA mrna RNA RNA mrna

160 2 mrna 78 --

161 (proteome) (proteomics) v

162 3 80 v 1 DNA [DNA chips] v 2 v 3 MS

163 4 Bioinformatics 81 v 21 (Genomics) (Proteomics)

164 82 v DNA GenBank EMBL v SWISS-PROT PIR MIPS v PDB v SCOP CATH

165 83 v 1 Alignment BLAST FASTA v 2 v Homology Threading 30

166 84 v v v 4 ( ) DNA 3~5% DNA DNA DNA 6 DNA

167 85 v v v 7 Contigs 500 Contigs EST NP

168 86 DNA

169 87 The Thank End You

hm 2 63% 980hm 2 19% 680hm 2

hm 2 63% 980hm 2 19% 680hm 2 1 2 b 3 4 Smith DNA DNA Berg l DNA DNA DNA Cohen DNA E. cloi 5 6 1996 1997 1998 1999 2000 2001 2002 1996 170 1997 1100 1998 2780 1999 3990 2000 4420 2001 5260 2002 5000 7 8 2001 100 3330hm 2 63% 980hm