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
Chapter 5. Molecular Genetic Techniques
CHAPTER 5
Molecular Genetic Techniques
© 2008 W. H. Freeman and Company
Teaching Plan- Chapter 5
April 9 , 2015
Activity
Scheduled Action/ timing
Class Test Chapter 4
10 minutes
Presentation by Vicky
5 mins
Lecture Continued Chapter 5
Video-- Expression Cloning of Receptors
Plasmid Cloning(Section 5.2)
5 minutes
4 minutes
Assignment- Review chapter/section
covered in class,
Quiz in next class, 5-10 minutes
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
Page no. 165- 175
5.1 166 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
P.N. 168
FIGURE 5-3
Comparision of
mitosis and
meiosis
P.N. 169 FIGURE 5-4(a)
Segregation patterns of
dominant and recessive
mutations in crosses
between true-breeding
strains of diploid
organisms
P.N. 169 FIGURE 5-4(b)
Segregation patterns of
dominant and recessive
mutations in crosses
between true-breeding
strains of diploid
organisms
Teaching Plan- Chapter 5
April 9 , 2015
Activity
Scheduled Action/ timing
Class Test Chapter 4
10 minutes
Presentation by Vicky
5 mins
Lecture Continued Chapter 5
Video-- Expression Cloning of Receptors
Plasmid Cloning(Section 5.2)
5 minutes
4 minutes
Assignment- Review chapter/section
covered in class,
Quiz in next class, 5-10 minutes
Conditional Mutations
Conditional Mutations
Can Be Used to Study
Essential Genes in Yeast
P.N. 170 FIGURE 5-6(a)
Haploid yeasts carrying
temperature-sensitive
lethal mutations are
maintained at permissive
temperature and analyzed
at nonpermissive
temperature
P.No 170 FIGURE 5-6(b)
Haploid yeasts carrying
temperature-sensitive
lethal mutations are
maintained at permissive
temperature and analyzed
at nonpermissive
temperature
P.No. 172 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
P.No 173 FIGURE 5-8(a)
Analysis of double mutants
often can order the steps in
biosynthetic or signaling
pathways
P.No 173 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
P.No 173 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
P.No 174 FIGURE 5-10(a)
Recombination during meiosis
can be used to map the
position of genes
Linkage Analysis
P.No 174 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
linkage
遗传标记
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
Transgenic
转基因
RNA interference (RNAi)
Micro RNAs (miRNAs)
Other key words in p212
Review the Concepts in p212
Analyze the Data in p213
小RNA