《基因诊断与基因治疗概论课件Gene_D_T_08-3》由会员分享，可在线阅读，更多相关《基因诊断与基因治疗概论课件Gene_D_T_08-3（53页珍藏版）》请在金锄头文库上搜索。
1、基因诊断与基因治疗概论（3）An Introduction toFan Leming Athrosclerosis Research Center Nanjing Medical UniversityCONTENTS内 容 IntroductionIntroduction Why do gene diagnosis and therapy Why do gene diagnosis and therapy Conception What is gene diagnosis and therapygene diagnosis and therapy Methodology How do gene
2、 diagnosis and therapydo gene diagnosis and therapy Clinical Applications IntroductionIntroduction Why do gene diagnosis and therapy Why do gene diagnosis and therapy Conception What is gene diagnosis and therapygene diagnosis and therapy Methodology How do gene diagnosis and therapydo gene diagnosi
3、s and therapy Clinical Applications How do gene diagnosis and therapydo gene diagnosis and therapy Methodology方法学：基因诊断Disease is related not only to the structure of gene but also to the transcription or translation levelGene diagnosis may put in practice by:1. DNA diagnosis 2. RNA diagnosis3. Fluor
4、escence in situ Hybridization (FISH)4. Gene chipto analyze the structure of geneto measure the expression of geneto detect deletion of large fragmentto detect multiple mutation simultaneously基因诊断常用方法Disease is related not only to the structure of gene but also to the transcription or translation lev
5、elGene diagnosis may put in practice by:1. DNA diagnosis 2. RNA diagnosis3. Fluorescence in situ Hybridization (FISH)4. Gene chip基因诊断常用方法Gene diagnosisGene diagnosis1 1The abnormal structure of gene may be detected at DNA level directly. If the pathogenic gene or its structure and the details of mut
6、ation is uncertain, it is necessary to judge the existence of pathogenic gene indirectly by means of genetic markers. Such means is called Linkage analyses. The infectious disease may be diagnosed by detecting the exogenous DNA of pathogen.1. DNA DiagnosisDNA诊断w Direct detection of known gene mutati
7、onRestriction fragment analysis, etc.The abnormal structure of gene may be detected at DNA level directly. If the pathogenic gene or its structure and the details of mutation is uncertain, it is necessary to judge the existence of pathogenic gene indirectly by means of genetic markers. Such means is
8、 called Linkage analyses. The infectious disease may be diagnosed by detecting the exogenous DNA of pathogen.w Linkage Analysis Restriction fragment length polymorphism1. DNA DiagnosisDNA诊断The abnormal structure of gene may be detected at DNA level directly. If the pathogenic gene or its structure a
9、nd the details of mutation is uncertain, it is necessary to judge the existence of pathogenic gene indirectly by means of genetic markers. Such means is called Linkage analyses. The infectious disease may be diagnosed by detecting the exogenous DNA of pathogen.1. DNA DiagnosisDNA诊断w Detection of exo
10、genous DNA1. DNA DiagnosisDNA诊断Direct detection of gene mutation1、Restriction fragment analysis2、Oligonucleotide dot blot analysis3、Amplification refractory mutation system 4、Single-strand conformation polymorphism5、 DNA sequencingDetection of exogenous DNALinkage Analysis Restriction fragment lengt
11、h polymorphism1. DNA DiagnosisDNA诊断Direct detection of known gene mutation1、Restriction fragment analysis2、Oligonucleotide dot blot analysis3、Amplification refractory mutation system 4、Single-strand conformation polymorphism5、 DNA sequencingDetection of exogenous DNALinkage Analysis Restriction frag
12、ment length polymorphismTo judge the existence of known mutation.Be applicable to cases in which the pathogenic gene is certain and its sequence is complete or partial known.（1）Direct detection of gene mutationCommon techniques：1) Restriction fragment analysis Be applicable to larger fragment mutati
13、on or point mutation located in restrictive enzyme site 2) Oligonucleotide dot blot hybridization analysis Be applicable to various point mutation, particularly outside of the restrictive enzyme site. 突变的直接检测1) Restrict fragment analysis Larger fragment deletionPoint mutation located in restrictive
14、enzyme site 限制性片断分析1.15kb 1.35kbbA/ bSbAbS1.35kbbS CCT GTG GAG1.15kb 0.2bA CCT GAG GAGMstII CCTNAGGSickle Cell Anaemia: Point mutation in coden 6 of b-globin GAG(Glu) GTG(Val)Example 1. Point mutation which leads to loss or gain of restrictive enzyme site 限制性片断分析：例 1HomoHeteroNormal5.2kb4.6kbExample
15、 2. Larger fragment mutation which leads to restrictive enzyme site shift b-Thalassemia ：0.6kb deletion at 3end of b-globin| | BgI II BgI II4.6kb| |BgI II BgI II 5.2kb限制性片断分析：例 2a a 14kbExample 3. Gene deletiona-Thalassemia ： various number (1-4) of b-globin gene deletiona10kb14kb10kbaa/aa aa/- aa/a- a-/- -/-BamHI BamHI限制性片断分析：例 3Combination of alleles1/1 1/2 1/3 1/4 2/2 2/3 2/43/3 3/4 4/4Example 4. Repeat sequenceSouthern blotAllele 1Allele 4Allele 3Allele 2限制性片断分析：例 4CCGGC CCGTCHp
