Molecular Cell Biology

1. 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. Quiz Analyzing Data Chapter 5 2. Chapter 5.4-5.5: Key Figures 4. Answer Questions

2. KEY CONCEPTS OF SECTION 5.4 Identifying and Locating Human Disease Genes(p204)

3. 5.4 Identifying and Locating Human Disease Genes

4. Further Analysis Is Needed to Locate a Disease Gene in Cloned DNA FIGURE 5-38 The relationship between the genetic and physical maps of a human chromosome

5. 6.2 Chromosome Organization of Genes and Noncoding DNA • Genomes of Many Organisms Contain Much Nonfunctional DNA • Most Simple-Sequence DNAs Are Concentrated in Specific Chromosomal Locations • DNA Finger Printing Depends on Differences in Length of Simple Sequence Repeat (SSR) DNAs -Also called STR (Short Tandem Repeat) DNA Profiling)

6. Generation of microsatellite repeats FIGURE 6-5(a) Generation of microsatellite repeats by backward slippage of the nascent daughter strand during DNA replication Most Simple-Sequence DNAs Are Concentrated in Specific Chromosomal Locations

7. Generation of microsatellite repeats FIGURE 6-5(b) Generation of microsatellite repeats by backward slippage of the nascent daughter strand during DNA replication

8. Hybridization of Simple Sequence DNA FIGURE 6-6 Simple-sequence DNA is localized at the centromere in mouse chromosome

9. STR DNA Fingerprint in Practice

10. DNA profiling by SSR (STR) Technique DNA Fingerprinting Depends on Differences in Length of Simple-Sequence DNAs FIGURE 6-7 DNA fingerprinting is used to identify individuals in paternity cases and criminal intestigations

11. KEY CONCEPTS OF SECTION 5.5 • Inactivating the Function of Specific Genes in Eukaryotes(p211) • Knockout-by Targeting Plasmid Vector • Conditional KO-by Lox-Cre Recombinase System • Knockdown-by SiRNA • Transgenic-byDominant Negative Protein cDNA

12. Specific Genes Can Be Permanently Inactivated in the Germ Line of Mice EXPERIMENTAL FIGURE 5-40(a) Isolation of mouse ES cells with a gene-targeted disruption is the first stage in production of knockout mice

13. EXPERIMENTAL FIGURE 5-40(b) Isolation of mouse ES cells with a gene-targeted disruption is the first stage in production of knockout mice

14. EXPERIMENTAL FIGURE 5-41 ES cells heterozygous for a disrupted gene are used to produce gene-targeted knockout mice

15. Conditional Knockout: To Study Embryonic Lethal Essential Gene KO

16. Somatic Cell Recombination Can Inactivate Genes in Specific Tissues EXPERIMENTAL FIGURE 5-42 The loxP-Cre recombination system can knock out genes in specific cell types

17. Dominant-Negative Alleles Can Functionally Inhibit Some Genes EXPERIMENTAL FIGURE 5-43 Transgenic mice are produced by random integration of a foreign gene into the mouse germ

18. FIGURE 5-44 Inactivation of the function of a wild-type GTPase by the action of a dominant-negative mutant allele

19. RNA Interference Causes Gene Inactivation by Destroying the Corresponding mRNA EXPERIMENTAL FIGURE 5-45 RNA interference (RNAi) can functionally inactivate genes in C.elegans and other organisms

20. Discussion: • Answer Chapter 5 Questions • Homework: Review Chapter 5 • Key Terms (p212) • Concepts p212 (will be tested in Final) • Analyzing the data p213-214 • (These will be tested in Final)

21. KEYWORDS