Teaching Plan- Chapter 5
March 19, 2015
Activity
Scheduled Action/ timing
Teaching Assistant Introduction Brief up
A short quiz Demo
Lecture by Prof Li. Chapter 3
2 minutes
10 minutes
Purifying, detecting, and Characterizing
Proteins (p92)
Videos- SDS Gel filtration
3 minutes
Lecture Continued
Video– Gel, Affinity and ion exchange
chromatography
8 minutes
Assignment- Review chapter/section
covered in class,
Quiz in next class, 5-10 minutes
Class test Chapter 3 (Analyze the data )
Scheduled on 24 March, 2013
Important Information
• Evaluation :
 60 % Class Performance






Attendance
Class interaction
Class tests
Group Performance
Presentation skills
Quiz
 40 % Final Exams



Tests are important so that we can grade you.
Easy scoring
Better understanding


Its very important to let us know your concerns or queries regarding the class and the subject
Suggestions are most welcome and appreciated !!!
Lodish Website details
• http://bcs.whfreeman.com/lodish7e
3.6 Purifying, detecting, and Characterizing Proteins (p92)
Differential
Centrifugation
Rate-Zonal Centrifugation
EXPERIMENTAL FIGURE 3-34
Centrifugation techniques
separate paticles that differ in
mass or density
Electrophoresis
SDS-PAGE
EXPERIMENTAL FIGURE
3-35
SDS-polyactrylamide gel
electrophoresis(SDSPAGE) separates proteins
primarily on the basis of
their masses
2-D Gel Electrophoresis
Page 95 EXPERIMENTAL
FIGURE 3-36
Two-dimensional gel
electrophoresis
separates proteins on
the basis of charge and
mass
Page 95 EXPERIMENTAL
FIGURE 3-36
Two-dimensional gel
electrophoresis
separates proteins on
the basis of charge and
mass
Video
• SDS Gel Electrophoresis (Website )
• http://bcs.whfreeman.com/lodish7e/#800911
__811695__
Page 96 Liquid Chromatography
EXPERIMENTAL FIGURE
3-37(a)
Three commonly used
liquid chromatographic
techniques separate
proteins on the basis of
mass, charge, or affinity
for a specific binding
partner
Page 96 EXPERIMENTAL FIGURE 3-37 (b)
Three commonly used liquid chromatographic
techniques separate proteins on the basis of
mass, charge, or affinity for a specific binding
partner
EXPERIMENTAL FIGURE
3-37(c)
Three commonly used
liquid chromatographic
techniques separate
proteins on the basis of
mass, charge, or affinity
for a specific binding
partner
Video
• Gel Filtration Chromatography
• Ion Exchange Chromatography
• Affinity Chromatography
Page 99 Western Blotting/Immunoblotting
EXPERIMENTAL FIGURE 3-38
Western blotting (immunoblotting)
combines several techniques to
resolve and detect a specific protein
Video
• Western Blot
http://bcs.whfreeman.com/lodish7e/#800911__
811695__
Radio Labeling
Radioisotopes are indispensable tools for detecting biological molecules
Page 101 Pulse-Chase labeling
Labeling Experiments and Detection of Radiolabeled Molecules
EXPERIMENTAL FIGURE 3-39
Pulse-chase experiments can
track the pathway of protein
modification or movement
within cells
Page 101 Mass Spectrometry
Mass Spectrometry Can Determine the Mass and Sequence of Proteins
EXPERIMENTAL FIGURE 3-40
Molecular mass can be determined by matrixassisted laser desorption/ionization time-of –
flight (MALDI-TOF) mass spectrometry
Page 102 Electrospray Ionization –Ion Trap-Mass
EXPERIMENTAL FIGURE 3-41(a)
Molecular mass of proteins and
peptides can be determinded by
electrospray ionization ion-trap
mass specrometry
EXPERIMENTAL FIGURE 3-41(a)
Molecular mass of proteins and
peptides can be determinded by
electrospray ionization ion-trap
mass specrometry
Page 104 LC-MS/MS can determine protein sequence
Determine 3-D structure by X-ray Crystallography
EXPERIMENTAL FIGURE 3-42(a)
X-ray crystallography provides
diffraction data from which the
three-dimensional structure of a
protein can be determined
Analysis
EXPERIMENTAL FIGURE 3-42(a)
X-ray crystallography provides
diffraction data from which the
three-dimensional structure of a
protein can be determined
Page 106 3.7 Proteomics-Definition and Techniques
Advanced Techniques in
Mass Spectrometry Are
Critical to Proteomic
Analysis
EXPERIMENTAL FIGURE
3-43
LC-MS/MS is used to
identify the proteins in a
complex biological
sample
Page 107 Identify Proteins: Density Gradient Centrifugation and LC-MS/MS
EXPERIMENTAL FIGURE 3-44(a)
Density-gradient centrifugation
and LC-MS/MS can be used to
identify many of the proteins in
organilles
EXPERIMENTAL FIGURE 3-44(a)
Density-gradient centrifugation
and LC-MS/MS can be used to
identify many of the proteins in
organilles
KEY CONCEPTS OF SECTION 3.7
Proteomics
Key words in chapter 3.6 and 3.7
Isoelectric point(pI)
等电点
Liquid chromatography(LC) 液相色谱
Western blotting
Radioisotope
放射性同位素
Pulse-chase
脉冲追踪
Proteomics
蛋白质组学
Prepare Student Presentations
Select 1 topic
The principle and Application
1. Gel Electrophoresis
2. Microarray
3. Immunostaining
KEY WORDS
Deoxyribonucleic acid(DNA)
Ribonucleic acid(RNA)
Messenger RNA(mRNA)
Transfer RNA(tRNA)
Ribosomal RNA(rRNA)
Transcription
Gene expression
Purine
Pyrimidine
Phosphodiester bond
Double helix
Base pair
complementary
脱氧核糖核酸
核糖核酸
信使ＲＮＡ
转运ＲＮＡ
核糖体ＲＮＡ
转录
翻译
嘌呤
嘧啶
磷酸二酯键
双螺旋
碱基
互补
Denaturation
Nucleic acid hybridizaiton
Micro RNAs(miRNAs)
RNA polymerase
Upstream
Promoter
Transcription factor
Primary transcript
Genome
Operon
Precursor mRNAs(pre-mRNAs)
RNA splicing
Alternative splicing
Isoform
Anticodon
Extron
Intron
变性
核酸杂交
小ＲＮＡ
ＲＮＡ聚合酶
上游
启动子
转录因子
原始转录的RNA
基因组
操纵子
前体ｍＲＮＡ
ＲＮＡ剪接
可变剪接
异构体
反义密码子
外显子
内含子
Ribosome
Genetic code
Reading frame
Aminoacyl-tRNA
Initiation factors(IFs)
Elongation factors(EFs)
Release factors(RFs)
GTPase superfamily
Polyribosome
Helicase
Leading strand
Lagging strand
Okazaki fragment
Cyclin-dependent kinase
Recombination
核糖体
密码子
阅读框
氨酰ｔＲＮＡ
起始因子
延伸因子
释放因子
ＧＴＰ酶超家族
多核糖体
解旋酶
前导链
后随链
冈崎片断
ＣＤＫ
重组
Point mutation
Meiosis
Crossing over
Virion
Bacteriophage
Capsid
Nucleocapsid
Envelope
Plaque assay
Lysogeny
Retrovirus
Reverse transcriptase
Provirus
Onco-gene
点突变
减数分裂
交换
病毒
噬菌体
衣壳
核衣壳
外壳
噬菌斑试验
溶原现象
反转录病毒
反转录酶
前病毒
致癌基因

Discussion:
Answer Questions:
1. How to Preview and Review
2. How to Present

Homework:
1.
2.
Review Chapter 4 Concepts p161 (will be tested in Final)
Analyzing the data p161-162
(These will be tested in Final)
3. Prepare quiz for Chapter 5
Next Tuesday –
Class test Chapter 3 questions and (Analyze the data )
Scheduled on 24 March, 2015