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1、regulation,分子生物学技术 （原理及应用）,课程总结,第一章-绪论，DNA和RNA结构 第二章-染色体，染色质和核小体(Chromosomes, chromatin, and the nucleosome） 第三章-DNA的复制(The replication of DNA） 第四章-DNA的突变和修复(The mutability and repair of DNA） 第五章-转录机制(Mechanisms of Transcription) 第六章 RNA剪接(RNA Splicing） 第七章 翻译-1，2, 3 第八章-遗传密码 (genetic code) 第九章-分子生物
2、学技术(Techniques of molecular biology) 第十章 原核生物基因表达调控(Regulation in prokaryotes) 第十一章 真核生物基因表达调控(Regulation in eukaryotes) 第十二章-DNA重组,第一章-绪论，DNA和RNA结构,分子生物学的含义 分子生物学的发展 分子生物学研究内容,What is Molecular biology?,Define in broadly: （广义定义） understand biological phenomena in molecular terms (difficult to disti
3、nguish from biochemistry),Define in restrictively: （狭义定义） the study of gene structure and their activities in molecular level（分子水平上研究基因的结构和功能）,Gene concept Gene structure Gene replication Gene expression Gene recombination Gene mutation,Molecular Biology of Gene （基因的分子生物学）,第二章-染色体，染色质和核小体(Chromosome
4、s, chromatin, and the nucleosome）,Chromosome sequence & diversity （染色体序列和多样性） The nucleosome（核小体） Higher-order chromatin structure （染色质的高级结构） Regulation of chromatin structure （染色质结构的调控）,Nucleosomes are the building blocks of chromosomes（核小体是染色体的结构单位）,在真核细胞中大多数DNA被包装进核小体 The nucleosome is composed o
5、f a core of eight histone proteins and the DNA (core DNA, 147 bp，核心DNA) wrapped around them（核小体由8个组蛋白所形成的核组成，DNA缠绕在组蛋白核上） The DNA between each nucleosome is called a linker DNA（连接DNA). Each eukaryote has a characteristic average linker DNA length (20-60 bp),Figure 7-18 DNA packaged into nucleosome,S
6、ix-fold DNA compaction,DNA盘绕在组蛋白核上 （H2A, H2B, H3，H4）,第三章-DNA的复制(The replication of DNA）,1. The Chemistry of DNA Synthesis(DNA合成的化学基础） 2. The Mechanism of DNA Polymerase（DNA聚合酶的作用机制） 3. The Specialization of DNA Polymerases（DNA聚合酶的特化） 4. The Replication Fork（复制叉） 5. DNA Synthesis at the Replication F
7、ork（复制叉上的DNA合成） 6. Initiation of DNA Replication（复制的起始） 7. Binding and Unwinding（结合和解旋） 8. Finishing Replication（复制的终止）,The Chemistry of DNA Synthesis: substrate, direction and energy. The Mechanism of DNA Polymerase: 1 polymerization mechanism, 2 different ways of discriminating substrates, 2 catal
8、ytic sites; 3 domains. The Specialization of DNA Polymerases The Replication Fork: the enzyme/proteins required to synthesize the leading and lagging strands. DNA Synthesis at the Replication Fork: Holoenzyme/trombone model to explain how the anti-parallel template strands are copied/replicated towa
9、rd the replication fork. Replisome/protein interaction.,Summary,Initiation of DNA Replication/binding and unwinding: the replicon model; initiation in bacteria; initiation control in eukaryotes-a link with cell cycle (pre-RC assembly and activation). Finishing Replication: Telomeres, telomerase,PCR（
10、聚合酶链式反应）,Polymerase chain reaction -An important technique based on DNA Polymerase,The polymerase chain reaction(PCR) is to used to amplify a sequence of DNA in vitro, using a pair of primers each complementary to one end of the the DNA target sequence.,复制理论,实验技术,Denaturation (变性): The target DNA (t
11、emplate) is separated into two stands by heating to 95 Primer annealing (退火): The temperature is reduced to around 55 to allow the primers to anneal. Polymerization (elongation, extension) (延伸): The temperature is increased to 72 for optimal polymerization step which uses dNTPs and requires Mg+.,The
12、 principle of PCR: Three different steps proceed in each PCR cycle.,Reverse transcriptase (RT)-PCR 逆转录PCR,AAA(A)n,5-Cap,mRNA,(dT)1218 primer,anneal,5-Cap,AAA(A)n,3,5,Reverse transcription,dNTP, RT,5-Cap,AAA(A)n,5,cDNA:mRNA hybrid,Regular PCR,第四章-DNA的突变和修复(The mutability and repair of DNA）,Replicatio
13、n errors and their repair (复制错误及其修复） DNA damage （DNA损伤） Repair of DNA damage（DNA损伤的修复）,复制中产生的错误以及它们是如何被修复的 自发产生或来自外界的攻击造成的各种损伤 细胞修复损伤的多种修复机制,生物在保持遗传物质忠实性方面的努力,复制中产生的错误以及它们是如何被修复的 自发产生或来自外界的攻击造成的各种损伤DNA损伤的各种类型 细胞修复DNA损伤的多种修复机制： 1. Direct reversal of DNA damage by photoreactivation (光活化作用) and alkyltr
14、ansferase (烷基转移酶)-直接将损伤逆转 2. 剪切修复系统（excision repair system)： 仅仅将受损的核苷酸去除 将包含损伤的一小段单链DNA去除 3. 重组修复系统（Recombination repair）：当DNA两条链都受损（断裂）时采用这种修复双链断裂修复（ Double-strand break repair） 4. 移损DNA合成（Translesion DNA synthesis）：DNA聚合酶的复制进程被受损碱基阻碍时采用的修复,1. Direct reversal of DNA damage by photoreactivation (光活化
15、作用) and alkyltransferase (烷基转移酶)-直接将损伤逆转 2. 剪切修复系统（excision repair system)：另一条未受损的链作为模板，以便DNA聚合酶重新掺入正确的核苷酸 仅仅将受损的核苷酸去除 将包含损伤的一小段单链DNA去除 3. 重组修复系统（Recombination repair）：当DNA两条链都受损（断裂）时采用这种修复双链断裂修复（ Double-strand break repair） 4. 移损DNA合成（Translesion DNA synthesis）：DNA聚合酶的复制进程被受损碱基阻碍时采用的修复,Mechanisms t
16、o repair a damage （DNA损伤的修复系统）,第五章-转录机制(Mechanisms of Transcription),RNA polymerase and the transcription cycle（RNA聚合酶和转录周期） The transcription cycle in bacteria（细菌的转录周期） Transcription in eukaryotes（真核生物的转录）,RNA polymerases (RNAP, 真核和原核的异同) and transcription cycle (Initiation, elongation and termination) Transcription cycle in bacteria: -Initiation: (1) The feature of s70 promoters. (2) Promoter binding by s70 transcription factor (recognition mechanism). (3) T
