Structure & Organization of Genes The Key Laboratory of Molecular Medicine Ministry of Education Weihh (weiyh@fudan.edu.cn) 2010.09
2 1. (a Unit of Inheritace) RNA DNA 2. (Nature of gene) heredity heredity & variation variation
3 Concept of the gene
(two basic questions) 4 (central dogma) Structure Function
5 v 1 DNA v 2 & 1 2 v 3
6 DNA Composition and
7 1. 2. DNA 3. DNA 4. DNA 5. DNA 6. DNA
DNA RNA 8 ( ) A G C T =
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
DNA 10 A, G, C, T AGGACCTTGAAGGTTCCAAATTGG 01 DNA DNA
DNA 11
12
(base match) 13 A=T, C G
DNA 14
B-DNA 15 v 0.34nm 10 bp 3.4nm 34Å v DNA v C G CG C2
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 34Å
z-dna 18
DNA 19
20 DNA
21 DNA 174 DNA 5 -(T X G Y ) n (A X C Y ) n -3
22
DNA 23 5 -(T X G Y ) n (A X C Y ) n -3 1. DNA 2. 3. telomerase 1. Azzalin, CM; Lingner, J, 2007. Telomeric Repeat Containing RNA and RNA Surveillance Factors at Mammalian Chromosome Ends. Science, Vol. 318. no. 5851, pp. 798 801 2. Azzalin, CM; Lingner, J, 2004. Telomere wedding ends in divorce. Science, Vol. 304. no.5667, pp. 60-62
6 DNA 24 v denaturation denaturation v renaturation renaturation annealing annealing DNA
Polymerase Chain Reaction, PCR 25
26 Molecular hybridization
DNA 27 v v v Tm 50%DNA
28
29 GC AT
Tm 30 DNA Tm Tm v 1 DNA v 2 DNA G+C G+C v 3
DNA 31 1. DNA Tm25 55 55 65 65 2. DNA 3. DNA 4. 0.18M Na + 5.
32 Genome & Genome Organization
33 v 1 (gene) v 2 (genome) v 3 (genome organization)
34
35 1. 2. 3. 4. 5.
1.1 36 X174 Open Reading Fram, ORF A B
1.2 37
1.3 38 mrna X174 PD
39 λ
1.5 40
41 λ
42 v 1 v 2 Operon Promoter mrna 600 v 3 v 4 v 5 trna rrna RNA rrna trna
lac Operon 43 trp Operon
44
trna rrna 45
46 v 1 v 2 v 3 v 4 v 5
47
48 v 1 30% <3% v 2 70% 3%
49 v 25 500kb v 10 v 20
50 <3% 3~4 1. 2. 3. 4.
1 51
2 52, interrupted gene
53 DNA mrna 8 mrna 7 7
(intron) (exon) 54 v (intron) v (exon):
55
56
57 GT/AG v : 5 GT,3,3 AG AG 5 3 GT AG
3 58
59 simple eukaryotic transcription units mrna complex eukaryotic transcription units mrna,
1 60
2 mrna 61 ( RNA) mrna. a b T
62 v 1 v 2 Splicing Form v 3
63 v v
4 gene family 64 v 25% e.g. v 2. v
4 65 v 1. v 2. v 3. v 4.
4.1 66 : rrna trna
4.1.1 rrna 67 v 28S 18S 5.8SrRNA 13 14 15 21 22 v 5SrRNA 1
68 1 5SrRNA 2000
4.1.2 trna 69 v v v trna
4.1.3 70 DNA 27% 66% RNA 6% ( )
71
4.1.3 72 5
4.2 72 v GH CS PRL v
74 v v v v v
75 2 2 2 2 2 1
76 ε δ HS
77
4.3 78 v src 250 AA v DEAD 8 AA Asp-Glu-Ala-Asp
4.4
4.4.1 04 Fab Fc Fc
79 v 10 9 v 10 9 10 12 bp
4.4.2 05 v Ig combinatorial joining v V C D J (junctional flexibility) v (somatic hypermutation)
4.4.3 Ig 06 v v IgG IgA IgM IgD IgE Ig 2
07 2 22 14
08 v v
09 100~1000 20 9 250-1000 12 4
4.4.4 10
11 v v
K 12
13
14 V H ND H NJ H V
class switch 15
16
17 B 10 5 V-D-J B
5 pseudogene 18 duplication v v v v ( )
19
1 DNA 20 v GC v GC
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),
22 2 (minisatellite)dna: v DNA GC 10~15bp v DNA Southern blot ~1000 DNA v DNA
23 3 (microsatellite)dna : v 10~50kb 1 DNA STR ( ),.
2 24 DNA transposon DNA - cut-and-paste retrotransposon RNA DNA LTR LINE SINE
25 v 1 LTR (long terminal repeat) 20000 v 2 LINE (long interspersed nuclear elements ) mrna 6.1kb 3500 v 3 SINE (short interspersed nuclear elements ) LINE Alu
Alu 26 DNA 130bp 170bp 30-50
27
28 DNA polymorphism
29 v DNA 99.9% 0.1% 0.1% DNA v DNA DNA DNA 1 DNA
3.1 DNA 30 v RFLP v (TR) v (SNP)
3.1.1 Restriction Fragment Length Polymorphism, RFLP) 31 v RFLP DNA v DNA 10 v 1. 2. 3.
32
3.1.2 33 v ( ) ( VNTR) v ( ) (STR)
( ) VNTR) 34 v Variable Number of DNA 15~65bp 1 2 3
35 v DNA v
(STR) 36 v 1~8bp 2 3bp v DNA 2~4bp v
37 10 CA 12 CA 9 CA 10 CA 9 CA
3.1.3 Single Nucleotide Polymorphism,SNP) 38 SNP v SNP v ACTGGCATTAACCCATT AGTGGCTTTAACCCATT v 1000bp SNP
SNP 39 v 300 1000 / v v SNP SNP
40 4 5-10%
SNP 41 v SNP DNA v 300~1000 SNP v HapMap
HapMap 42 v. v (Haplotype) 100 300 SNP. v SNP (Haplotype Map, HapMap)
HapMap v v v v,
3.2 DNA 43 v 1 v 2 I II
44
45 X X 1/1200 1/2500 80% 20%
X FMR1 46 FMR1(Fragile X Mental Retardation-1) X FMR1 5 - (CGG)n v v v 1 n 7~60 30 2 n 60~230 FMR1 3 n 230 FMR1
e.g. 47 1 2 3 4 5 6 100 60
48 1 2 3 4 5 6 7
3.3.1 49 v RFLP Huntington s disease HD v 10 20 55 v
50 v IT15 4P6.3 HD v CAG 6 35 > 37 100 HD 17 v HD v HD
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
3.3.2 52 GAA Glu GUA Val
DNA DNA typing 53 v DNA DNA v RFLP STR VNTR SNP v 1. DNA 2. PCR STR/ VNTR 3. SNP
DNA 54 VNTR DNA
个体识别 鉴定罪犯 55
PCR-STR 56 STR STR STR 100 200bp DNA PCR DNA
57 v proto-oncogene oncogene akt ets H-ras, hst-1 v anti-oncogene Rb p53 APC v
(The Cancer Genome Atlas, TCGA) v DNA,, v
58 30 60 220 / 10 / 100 v 10 40 20 30
Pharmacogenomics) 59 v
60 v SNP SNP v
61 v Herceptin HER-2 25% 30% Herceptin
62 1 2 3 4 5 6
63 Part III Human HGP
64 DNA
65 4 1 2 3 4
1. genetic map 66 1 (cm,centimorgan)
67 v 1 ABO HLA 70 RFLP v 2 1985 minisatellite core VNTR 1989 microsatellite marker STR v 3 1996 MIT Lander ES SNP single nucleotide polymorphysm
2. map 68 DNA STS bp
69 v STS DNA contig DNA 1998 52,000 STS DNA
70 v DNA contig v v (sequence-tagged site, STS) DNA DNA v CpG Alu isochore v (cytogenetic map) v cdna/est
3. sequence map 71 DNA DNA DNA
72 1 (contig] 2 3 [sequencing]
4. gene map 73 cdna / ESTs expressed sequence tags 20
74 v v v
75
v 2.91Gbp 39000 14 v 300 v 26000 42% v 1/1250bp 140 99.99
v v v v v " " " " Y 200 SNP 30
76 (post-genome eva) v post-genome project
1 DNA 77 v gene targeting knock out knock in v gene silencing RNA mrna RNA RNA mrna RNA RNA mrna
2 mrna 78 --
(proteome) (proteomics) 79... v
3 80 v 1 DNA [DNA chips] v 2 v 3 MS
4 Bioinformatics 81 v 21 (Genomics) (Proteomics)
82 v DNA GenBank EMBL v SWISS-PROT PIR MIPS v PDB v SCOP CATH
83 v 1 Alignment BLAST FASTA v 2 v 3 2 3 Homology Threading 30
84 v v v 4 ( ). 20 5 DNA 3~5% DNA DNA DNA 6 DNA
85 v v v 7 Contigs 500 Contigs EST NP- 8 9 10 3
86 DNA
87 The Thank End You