大学分子生物学经典双语课件C2 GENE AND CHROMOSOME
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大学分子生物学经典双语课件C4-Gene_mutation_and_exchange资料
Meslson-Radding Model
2 Site-specific recombination
This involves the exchange of nonhomologous
but specific pieces of DNA and is mediated by
proteins (enzyme) that recognize specific DNA
or more genes conferring antibiotic resistance.
4.1.1.3 TnA family
TnA family is about 5000bp, which is much greater than the insertion sequence. The same as composite transposon, TnA family also carries the gene who is responsible for its own transposition and other gene such as resistance gene β -amine acyl enzyme (AmpR) . It has no IS, but there are terminal repeat sequences of about 37-38bp in the end.
4.4.2 Transposons in eukaryotes
4.4.2.1 Transposons in maize
Autonomous element
Nonautonomous element
（1）Ac-Ds system
11bp Ac
Ds
transposable element 是引起玉米糊粉层花斑不稳定
大学分子生物学经典双语课件
2.1.2.2 Conformation polymorphism of the double helix
Alternative doublehelical structures of DNA
Base Obliquity
helix rise per base pair
bp number per turn
biological activity changed (even lost)； viscosity decreased，粘度 solubility decreased,溶解度 Hyperchromicity: the absorbance of ssDNA is greater than that dsDNA.增色 concentration = 50μg/ml： dNTPs A260 = 1.60 S.S DNA A260 = 1.37 D.S DNA A260 = 1.0
2.1.3
Triplex DNA
1953, Watson & Crick proposed D.S DNA model and found many redundant hydrogen bonding donor and receptors along big grooves. 1957, Felsenfeld proposed T.S DNA concept
transferring the other dsDNA through the break.
Type I topoisomerase
Type II topoisomerase
Contents
1
2 3 4 5 6 7
Structure of DNA Denaturation, renaturation and hybridization
分子生物学全套课件96P课件97页PPT
Then what?
Amplification (PCR) +Separation
ROCHE Strategy
Emulsion PCR; 1 பைடு நூலகம்ragment/ bead
Primer
dNTP (each)
Polymerase
Template
Not clearly demonstrated
PCR amplification (Chain extension) Emulsion breaking Template dissociation
Around 100 million emPCR-prepared template beads are deposited onto a glass slide. On annealing of a universal primer, a library of 1,2-probes is added. Appropriate conditions enable selective hybridization and ligation of probes to complementary positions. The first (Y) and second (Z) positions of the 1,2-probes are designed as interrogation bases, such that the 16 dinucleotides are encoded by four dyes. Following four-colour imaging, the ligated 1,2probes are chemically cleaved to generate a 5′-PO4 group (P). The cycle of hybridization, ligation, imaging, and cleavage is repeated six more times. The extended primer is then stripped from the templates, and a second ligation round is performed with an n–1 primer, which resets the interrogation bases one position to the left. Interrogating each base twice improves the accuracy of the colour call. Seven ligation cycles ensue, followed by three more ligation rounds. A string of 35 data bits, encoded in colour space, are then aligned to a reference genome to decode the DNA sequence. Substitutions are the most common error type.
双语分子生物学ppt
主要参考教材
• PC Turner et al. Instant Notes in Molecular Biology (Second edition). BIOS Scientific Publishers Limited, 2000
• 分子生物学(第三版)导读版(精要速览系列) • 朱玉贤，李毅，郑晓峰编著。现代分子生物学（第3版） 高等教育出版
2.2 Prokaryotic and Eukaryotic chromosome structure
2.1 Properties of nucleic acid
2.1.1 Nucleic acid structure 2.1.2 Chemical & physical properties 2.1.3 Spectroscopic (光谱学) & thermal （热力学） properties 2.1.4 DNA supercoiling
– Glycerides (甘油酯), phospholipids(磷脂)，sphingolipids （鞘脂，如神经氨酰）
• Complex macromolecules
– Nucleoproteins(核蛋白), ribozyme（核糖体） – Glycoproteins – Proteoglycans (mucoproteins)蛋白多糖（粘蛋白） – Lipid-linked proteins – glycolipids
A, B and Z helices
• Z－DNA：它是左手双螺旋，与右手螺旋的不同是螺距延长 (4.5nm左右)，直径变窄(1.8nm)，每个螺旋含12个碱基对， 分子长链中磷原子不是平滑延伸而是锯齿形排列，有如“之” 字形一样，因此叫它Z构象，这一构象中的重复单位是二核 苷酸而不是单核苷酸；而且Z－DNA只有一个螺旋沟，它相当 于B构象中的小沟，它狭而深，大沟则不复存在。
分子生物学英文课件：molecular biotechnology
Analyze the recombinant plasmid and bacteriophage - screening DNA library.
(2) Northern blotting
❖ Similar to the southern blotting ❖ RNAs instead of DNAs ❖ No need of RE digestion ❖ Application
produce single strand DNA Transferring: transfer DNA to the NC. Immobilization: heating NC in 80℃ for 1～2 h
or uv crosslink (nylon membrane) Hybridization: NC is exposed to probe (biotin/
高等教育 > 生物学 > 分子生物学英文课件:molecular biotechnology molecularbiotechnology section molecularhybridization blottingtechnique heteroduplexfrom two complementary polynucleotide strands from different sources (dna dna;dna rna;rna molecularhybridization related conceptions/principles dnadenaturation dnarenaturation probedna rnafragment labeled radioisotope,biotin detectspecific nucleic acid sequences hybridization.?dna probe ?rna probe blottingtransfer (blot) biologic macromolecules separated fixthem nitrocellulose/nylonmembrane diffusion,electro-transferring vacuumabsorption, 1975,edwen southern blotting widelyused specificmacro- molecules (proteins, mrnas dnasequences) southernblotting genomic dna (from tissues re,separated gelelectro- phoresis nitrocellulosemembrane detectingspecific dna sequence labeledprobe. used qualitativelyanalyze genomic dna, recombinant plasmid, screening dna library. steps extraction:genomic dna from tissues digestion:cut separation:separate dna fragments denaturation:dna fragments treated p
(2) Northern blotting
❖ Similar to the southern blotting ❖ RNAs instead of DNAs ❖ No need of RE digestion ❖ Application
produce single strand DNA Transferring: transfer DNA to the NC. Immobilization: heating NC in 80℃ for 1～2 h
or uv crosslink (nylon membrane) Hybridization: NC is exposed to probe (biotin/
高等教育 > 生物学 > 分子生物学英文课件:molecular biotechnology molecularbiotechnology section molecularhybridization blottingtechnique heteroduplexfrom two complementary polynucleotide strands from different sources (dna dna;dna rna;rna molecularhybridization related conceptions/principles dnadenaturation dnarenaturation probedna rnafragment labeled radioisotope,biotin detectspecific nucleic acid sequences hybridization.?dna probe ?rna probe blottingtransfer (blot) biologic macromolecules separated fixthem nitrocellulose/nylonmembrane diffusion,electro-transferring vacuumabsorption, 1975,edwen southern blotting widelyused specificmacro- molecules (proteins, mrnas dnasequences) southernblotting genomic dna (from tissues re,separated gelelectro- phoresis nitrocellulosemembrane detectingspecific dna sequence labeledprobe. used qualitativelyanalyze genomic dna, recombinant plasmid, screening dna library. steps extraction:genomic dna from tissues digestion:cut separation:separate dna fragments denaturation:dna fragments treated p
分子生物学ppt课件
基因组大小(Mb)
0.58 1.83 4.20 4.60 13.50 12.50 466 165 97 2700 3000
基因数
470 1743 4100 4288 6034 4929 30000 13601 18424 30000 25000
染色体数*
无 无 无 无 16 16 21 4 6 20 23
包括：
结构基因组学
功能基因组学
三个亚领域.
比较基因组学
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一、病毒基因组 二、原核生物基因组 三、真核生物基因组
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一、病毒基因组
基因组（genome） 1个配（精子或卵子），1个单倍 体细胞或1个病毒所包含的全套遗传物质的总和。病毒核酸 或为DNA或为RNA，可以统称为病毒染色体。
完整的病毒颗粒具有蛋白质外壳，以保护病毒核酸不 受核酸酶的破坏，并能识别和侵袭特定的宿主。
分子生物学
Molecular Biology
1
What is Molecular Biology?
分子生物学是从分子水平研究生命现象、生命规律和生命本质 的学科。
核心内容是从分子水平研究基因和基因的活动，这些活动主要 通过核酸和蛋白质的活动来实现。
医学分子生物学主要研究人体生物大分子和大分子体系的结构、 功能、相互作用及其与疾病发生、发展的关系。
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三、基因的结构特点和分类
基因的结构
结构基因：编码区序列（coding region sequence ）
在细胞内表达为蛋白质或功能RNA的DNA序列
转录调控序列：非编码序列（non-coding sequence）
基因表达需要的调控区（regulatory region）序列， 包括启动子（promoter）、增强子（enhancer）等。
分子生物学(双语)10-Chromatin教学课件
– It is the basic level of organization of nucleosomes in chromatin.
• nonhistone – Any structural protein found in a chromosome except one of the histones.
DNA occupies most of the outer surface of the nucleosome
• footprinting – A technique for identifying the site on DNA bound by some protein by virtue of the protection of bonds in this region against attack by nucleases.
A protein protects a series of bonds against nuclease attack
FIGURE 05: Micrococcal nuclease initially cleaves between nucleosomes
The nucleosome is a cylinder with DNA organized into ~1 2/3 turns around the surface
The 30 nm fiber has a coiled structure
Photo courtesy of Barbara Hamkalo, University of California, Irvine.
DNA Is Organized in Arrays of Nucleosomes
• MNase (micrococcal nuclease) cleaves linker DNA and releases individual nucleosomes from chromatin.
• nonhistone – Any structural protein found in a chromosome except one of the histones.
DNA occupies most of the outer surface of the nucleosome
• footprinting – A technique for identifying the site on DNA bound by some protein by virtue of the protection of bonds in this region against attack by nucleases.
A protein protects a series of bonds against nuclease attack
FIGURE 05: Micrococcal nuclease initially cleaves between nucleosomes
The nucleosome is a cylinder with DNA organized into ~1 2/3 turns around the surface
The 30 nm fiber has a coiled structure
Photo courtesy of Barbara Hamkalo, University of California, Irvine.
DNA Is Organized in Arrays of Nucleosomes
• MNase (micrococcal nuclease) cleaves linker DNA and releases individual nucleosomes from chromatin.
分子生物学英文课件：Eukaryote gene and genome
Exon: segments of eukaryotic gene (or of the primary transcript of that gene) that are saved as part of a functional, mature mRNA, rRNA or tRNA molecules. Intron: segments of eukaryotic gene (or of the primary transcript of that gene) that are removed by splicing during RNA processing and are not included in the mature, functional mRNA, rRNA, or tRNA.
Repetitive sequences
高度重复序列（highly repetitive sequence） 中度重复序列（moderately repetitive sequence） 单拷贝序列（single copy sequence）或低度重复序列
（一）高度重复序列（highly repetitive sequence）
➢ Have unique structure elements
基因 -N
转录起点
+1
结构基因区
调控区
mRNA 5’
AUG
翻译起点
蛋白质 N
转录 翻译
转录终点
+N
UAA
3’
翻译终点
C
Gene expression: DNA￫mRNA￫Protein DNA ￫RNA
mRNA
mRNA
转录
tRNA
rRNA
Repetitive sequences
高度重复序列（highly repetitive sequence） 中度重复序列（moderately repetitive sequence） 单拷贝序列（single copy sequence）或低度重复序列
（一）高度重复序列（highly repetitive sequence）
➢ Have unique structure elements
基因 -N
转录起点
+1
结构基因区
调控区
mRNA 5’
AUG
翻译起点
蛋白质 N
转录 翻译
转录终点
+N
UAA
3’
翻译终点
C
Gene expression: DNA￫mRNA￫Protein DNA ￫RNA
mRNA
mRNA
转录
tRNA
rRNA