activityscheduled action/ timing teaching assistant introduction brief up a short quiz demo lecture...
TRANSCRIPT
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Activity Scheduled Action/ timing
Teaching Assistant Introduction Brief up A short quiz Demo Lecture by Prof Li. Chapter 3
2 minutes10 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
Teaching Plan- Chapter 5 March 19, 2015
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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 !!!
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3.6 Purifying, detecting, and Characterizing Proteins (p92)
Differential Centrifugation
EXPERIMENTAL FIGURE 3-34Centrifugation techniques separate paticles that differ in mass or density
Rate-Zonal Centrifugation
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Electrophoresis
SDS-PAGE
EXPERIMENTAL FIGURE 3-35SDS-polyactrylamide gel electrophoresis(SDS-PAGE) separates proteins primarily on the basis of their masses
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2-D Gel Electrophoresis
Page 95 EXPERIMENTAL FIGURE 3-36Two-dimensional gel electrophoresis separates proteins on the basis of charge and mass
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Page 95 EXPERIMENTAL FIGURE 3-36Two-dimensional gel electrophoresis separates proteins on the basis of charge and mass
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Video
• SDS Gel Electrophoresis (Website ) • http://bcs.whfreeman.com/lodish7e/
#800911__811695__
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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
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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
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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
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Video
• Gel Filtration Chromatography • Ion Exchange Chromatography• Affinity Chromatography
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Page 99 Western Blotting/Immunoblotting
EXPERIMENTAL FIGURE 3-38Western blotting (immunoblotting) combines several techniques to resolve and detect a specific protein
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Video
• Western Blot http://bcs.whfreeman.com/lodish7e/#800911__811695__
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Radio Labeling
Radioisotopes are indispensable tools for detecting biological molecules
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Page 101 Pulse-Chase labeling
EXPERIMENTAL FIGURE 3-39Pulse-chase experiments can track the pathway of protein modification or movement within cells
Labeling Experiments and Detection of Radiolabeled Molecules
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Page 101 Mass Spectrometry
EXPERIMENTAL FIGURE 3-40Molecular mass can be determined by matrix-assisted laser desorption/ionization time-of –flight (MALDI-TOF) mass spectrometry
Mass Spectrometry Can Determine the Mass and Sequence of Proteins
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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
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EXPERIMENTAL FIGURE 3-41(a)Molecular mass of proteins and peptides can be determinded by electrospray ionization ion-trap mass specrometry
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Page 104 LC-MS/MS can determine protein sequence
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Determine 3-D structure by X-ray Crystallography
Analysis
EXPERIMENTAL FIGURE 3-42(a)X-ray crystallography provides diffraction data from which the three-dimensional structure of a protein can be determined
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EXPERIMENTAL FIGURE 3-42(a)X-ray crystallography provides diffraction data from which the three-dimensional structure of a protein can be determined
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Page 106 3.7 Proteomics-Definition and Techniques
EXPERIMENTAL FIGURE 3-43LC-MS/MS is used to identify the proteins in a complex biological sample
Advanced Techniques in Mass Spectrometry Are Critical to Proteomic Analysis
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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
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EXPERIMENTAL FIGURE 3-44(a)Density-gradient centrifugation and LC-MS/MS can be used to identify many of the proteins in organilles
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KEY CONCEPTS OF SECTION 3.7Proteomics
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Key words in chapter 3.6 and 3.7
Isoelectric point(pI) 等电点Liquid chromatography(LC) 液相色谱Western blottingRadioisotope 放射性同位素Pulse-chase 脉冲追踪Proteomics 蛋白质组学
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1. Gel Electrophoresis
2. Microarray
3. Immunostaining
The principle and Application
Prepare Student PresentationsSelect 1 topic
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KEY WORDS
Deoxyribonucleic acid(DNA)Ribonucleic acid(RNA)Messenger RNA(mRNA)Transfer RNA(tRNA)Ribosomal RNA(rRNA)TranscriptionGene expressionPurinePyrimidinePhosphodiester bondDouble helixBase paircomplementary
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
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DenaturationNucleic acid hybridizaitonMicro RNAs(miRNAs)RNA polymeraseUpstreamPromoterTranscription factorPrimary transcriptGenomeOperonPrecursor mRNAs(pre-mRNAs)RNA splicingAlternative splicingIsoformAnticodonExtronIntron
变性核酸杂交小RNARNA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
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RibosomeGenetic codeReading frameAminoacyl-tRNAInitiation factors(IFs)Elongation factors(EFs)Release factors(RFs)GTPase superfamilyPolyribosomeHelicaseLeading strandLagging strandOkazaki fragmentCyclin-dependent kinaseRecombination
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
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Point mutationMeiosisCrossing overVirionBacteriophageCapsidNucleocapsidEnvelopePlaque assayLysogenyRetrovirusReverse transcriptaseProvirusOnco-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
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Discussion: Answer Questions:
1. How to Preview and Review 2. How to Present
Homework:
1. Review Chapter 4 Concepts p161 (will be tested in Final)2. 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