Molecular Cell Biology Professor Dawei Li daweili@sjtu.edu.cn 3420-4744 Textbook: MOLECULAR CELL BIOLOGY 6th Ed Lodish • Berk • Kaiser • Krieger • Scott • Bretscher •Ploegh • Matsudaira Part 2. Genetics and Molecular Biology 1. Review Chapter 4 2. Student Presentations 3. Chapter 5: Key Figures 4. Answer Questions

Analyze the Data: Chapter 3 Answer Keys Proteins 3, 5, 6, and 7 do not change in response to the drug. Protein 1 declines in response to the drug. Proteins 2 and 4 are induced in response to the drug. Proteins 2, 3 and 6 are phosphoproteins; the others are not. Protein 2 is a phosphoprotein normally present in cells whose level increases in response to the drug. Protein 3 is not a phosphoprotein in control cells and is phosphorylated in response to the drug. Protein 6 is a phosphoprotein whose level does not change in response to the drug. Proteins 1 and 6 are strictly nuclear proteins. Proteins 2, 3, and 4 are strictly cytoplasmic proteins. Protein 5 is present in the nucleus and cytoplasm. Protein 7 is a cytoplasmic protein that migrates to the nucleus in response to drug treatment.

d. Protein 1 is a nuclear protein whose level declines in response to drug treatment. Protein 2 is a cytoplasmic phosphoprotein whose level increases in response to drug treatment. Protein 3 is a cytoplasmic phosphoprotein whose level does not change in response to drug treatment. Protein 4 is a cytoplasmic protein whose level increases in response to drug treatment. Protein 5 is a nuclear and cytoplasmic protein whose level does not change in response to drug treatment. Protein 6 is a nuclear phosphoprotein whose level does not change in response to drug treatment. Protein 7 is a cytoplasmic protein which migrates from the cytoplasm to the nucleus, in response to drug treatment.

Viral Capsids Are Regular Arrays of One or a Few Types of Protein Review 4.7 Viruses Viral Capsids Are Regular Arrays of One or a Few Types of Protein FIGURE 4-44 Virion structures

Viruses Can Be Cloned and Counted in Plaque Assays EXPERIMENTAL FIGURE 4-45(a) The plaque assay determines the number of infectious particles in a viral suspension

EXPERIMENTAL FIGURE 4-45(b) The plaque assay determines the number of infectious particles in a viral suspension

Lytic Viral Growth Cycles Lead to the Death of Host Cells FIGURE 4-46 Lytic replication cycle of a nonenveloped bacterial virus

FIGURE 4-47 Lytic replication cycle of an enveloped, animal virus

EXPERIMENTAL FIGURE 4-48 Progeny virions are released by budding

Viral DNA Is Integrated into the Host-Cell Genome in Some Nonlytic Viral Growth Cycles FIGURE 4-49 Retroviral life sycle

© 2008 W. H. Freeman and Company CHAPTER 5 Molecular Genetic Techniques © 2008 W. H. Freeman and Company

OUTLINE 5.1 Genetic Analysis of Mutations to Identify and Study Genes 5.2 DNA cloning and Characterization 5.3 Using Cloned DNA Fragments to Study Gene Expression 5.4 Identifying and Locating Human Disease Genes 5.5 Inactivating the Function of Specific Genes in Eukaryotes

5.1 Genetic Analysis of Mutations to Identify and Study Genes FIGURE 5-1 Overview of two strategies for relating the function,location, and structure of gene products

Segregation of Mutations in Breeding Experiments Reveals Their Dominance or Recessivity FIGURE 5-3 Comparision of mitosis and meiosis

FIGURE 5-4(a) Segregation patterns of dominant and recessive mutations in crosses between true-breeding strains of diploid organisms

FIGURE 5-4(b) Segregation patterns of dominant and recessive mutations in crosses between true-breeding strains of diploid organisms

Conditional Mutations Conditional Mutations Can Be Used to Study Essential Genes in Yeast FIGURE 5-6(a) Haploid yeasts carrying temperature-sensitive lethal mutations are maintained at permissive temperature and analyzed at nonpermissive temperature

FIGURE 5-6(b) Haploid yeasts carrying temperature-sensitive lethal mutations are maintained at permissive temperature and analyzed at nonpermissive temperature

Complementation Analysis Complementation Tests Determine Whether Different Recessive Mutations Are in the Same Gene FIGURE 5-7 Complementation analysis determines whether recessive mutations are in the same or different genes

Double Mutations Are Useful in Assessing the Order in Which Proteins Function FIGURE 5-8(a) Analysis of double mutants often can order the steps in biosynthetic or signaling pathways

FIGURE 5-8(b) Analysis of double mutants often can order the steps in biosynthetic or signaling pathways

Genetic Suppression and Synthetic Lethality Can Reveal Interacting or Redundant Proteins FIGURE 5-9 Mutations that result in genetic suppression or synthetic lethality reveal interacting or redundant proteins

Linkage Analysis Genes Can Be Identified by Their Map Position on the Chromosome FIGURE 5-10(a) Recombination during meiosis can be used to map the position of genes

Linkage Analysis FIGURE 5-10(b) Recombination during meiosis can be used to map the position of genes

KEY WORDS Allete 等位基因 Mutation 突变 Mutagen Genotype 基因型 Wild type 野生型 Phenotype 表型 Haploid 单倍体 Diploid 二倍体 Heterozygous 杂合子 Homozygous 纯合子 Recessive 隐性 Dominant 显性 Point mutation 点突变 Gemete Mitosis 有丝分裂

Germ cells Meiosis 减数分裂 Homolog 纯合子 Homologous chromosome 同源染色体 Segregation Type a Temperature-sensitive mutation 温度敏感型突变株 Cell cycle 细胞周期 Genetic complementation Suppressor mutation Synthetic lethal mutation Genetic mapping Recombination 重组 Crossing over Locus Genetic marker 遗传标记 linkage

DNA cloning 基因克隆 Recombinant DNA 重组DNA Restriction enzyme 限制性内切酶 DNA ligase DNA连接酶 Restriction fragment 限制片段 Okazaki fragment 冈崎片段 Transformation 转化 DNA library DNA文库 Complementary DNAs (cDNAs) Reverse transcriptase 反转录酶 Probe 探针 Hybridization 杂交 Functional complementation Shuttle vector 穿梭质粒 Electrophoresis 电泳 Polymerase chain reaction (PCR) 聚合酶链式反应 Mobile DNA element

Southern Blotting Northern blotting Hybridization 杂交 DNA microarray DNA微阵列 Expression vector 表达载体 Promoter 启动子 Transfection 转染 Epitope 抗原决定簇 Autosome DNA-based molecular marker Restriction fragment length polymorphisms(RFLP) 限制片段长度多态性 Germ line 细胞系 Gene knockout 基因敲除 Embryonic stem cells 胚胎干细胞 Chimeras Dominant-negative mutation 显隐性突变 transgene 转基因

Review the Concepts in p212 Analyze the Data in p213 Transgenic 转基因 RNA interference (RNAi) Micro RNAs (miRNAs) 小RNA Other key words in p212 Review the Concepts in p212 Analyze the Data in p213