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BiochemistryChemicaloflife

2023/2/182Studyoflifeessentialityinmoleculelevels

从分子水平研究生命现象本质lifeessentiality

(生命的基本特征)metabolism(新陈代谢)genetics(遗传)growth(生长)development(发育)procreation(生殖)2023/2/183Biochemistrydiscipline(生物化学学科)

Origin(起源)chemistry、physiology、medicine、etc.acrossdiscipline(交叉学科)growthtop(生长点)Researchmethods(方法)chemical、physical、biological、etc.Objective(对象)biomolecule(生物分子)biomacromolecule(生物大分子)biomicromolecule(生物小分子)2023/2/184Biomacromolecules(生物大分子)Definitionmacromoleculemadeupbyorganicmicromolecule(componentmolecule,构件分子)specialconformation(构象)/configuration(构型)importantfunctionsMainbiomacromoleculeproteins(蛋白质)nucleicacids(核酸)lipids(脂类)polysaccharides(多糖)2023/2/185Biomicromolecules(生物小分子)organicandinorganicmicro/minimoleculeswithactivityinthecell(细胞内具有活性的小分子有机物和无机物)coenzymesandprostheticgroups(辅酶、辅基)vitamins(维生素)hormone(激素)exitatoryaminoacids(兴奋性氨基酸,Glu谷氨酸)freenucleicacids(游离核苷酸)microRNA(微小RNA),siRNA(干扰小RNA)ATP、ADP、AMP(腺苷酸)NO(一氧化氮)2023/2/186Maincontent(主要内容)Structureandfunctionofbiomolecules

生物分子的结构与功能(静态生物化学)Metabolismandregulationofbiomolecules

生物分子的代谢与调节(动态生物化学)Transmissionandexpressionofgenemessenger

生物遗传信息的传递与表达(机能生物化学,分子生物学)Signaltransductionofcellanddisorder

细胞信号转导(传导)与失常(机能生物化学,分子生物学)2023/2/187Maincontent(主要内容)Structureandfunctionofbiomolecules

生物分子的结构与功能Metabolismandregulationofbiomolecules

生物分子的代谢与调节Transmissionandexpressionofgenemessenger

生物遗传信息的传递与表达Signaltransductionofcellanddisorder

细胞信号转导(传导)与失常2023/2/188Hb2023/2/189Maincontent(主要内容)Structureandfunctionofbiomolecules

生物分子的结构与功能Metabolismandregulationofbiomolecules

生物分子的代谢与调节Transmissionandexpressionofgenemessenger

生物遗传信息的传递与表达Signaltransductionofcellanddisorder

细胞信号转导(传导)与失常2023/2/1810Glucosemetabolism

葡萄糖代谢Glucose+O2--->CO2+H2O+ATP

2023/2/1811Maincontent(主要内容)Structureandfunctionofbiomolecules

生物分子的结构与功能Metabolismandregulationofbiomolecules

生物分子的代谢与调节Transmissionandexpressionofgenemessenger

生物遗传信息的传递与表达Signaltransductionofcellanddisorder

细胞信号转导(传导)与失常2023/2/1812DNAReplication复制RNAReplicationReverseTranscription逆转录ProteinTranslation翻译Transcription转录Centraldogma(中心法则)2023/2/1813Maincontent(主要内容)Structureandfunctionofbiomolecules

生物分子的结构与功能Metabolismandregulationofbiomolecules

生物分子的代谢与调节Transmissionandexpressionofgenemessenger

生物遗传信息的传递与表达Signaltransductionofcellanddisorder

细胞信号转导(传导)与失常

2023/2/1814Signaltransduction(信号转导)signalmolecule(信号分子)hormone(激素)ligand(配体)

forexample:growthfactor(生长因子)receptor(受体)secondmessengercAMP、DG、IP3、Ca2+etc.signalingtransductionpathway

EGFFGFIGFPDGFTGF2023/2/18152023/2/18162023/2/1817Historyanddevelopmentancientlybiochemistry(古代生物化学)modernbiochemistry(现代生物化学)1770——1903prepare(静态生物化学)(动态生物化学)

(机能生物化学)1903——1953——现在1903Neuberg:Biochemistry(Establishedin1903)

1926Sumner:Enzymecanbecrystallized1937Krebs:Citricacidcycle1944Avery:Pneumonococcustransformationtest1950Pauling:α-helixofprotein2023/2/1818Historyanddevelopmentancientlybiochemistry(古代生物化学)modernbiochemistry(现代生物化学)1770——1903prepare1903——1953——现在1903Neuberg:Biochemistry(Establishedin1903)

1926Sumner:Enzymecanbecrystallized(结晶尿酶)1937Krebs:Citricacidcycle1944Avery:Pneumonococcustransformationtest1950Pauling:α-helixofprotein2023/2/1819Importantachievement

Sumner:Enzymecanbecrystallized2023/2/1820Historyanddevelopmentancientlybiochemistry(古代生物化学)modernbiochemistry(现代生物化学)1770——1903prepare1903——1953——现在1903Neuberg:Biochemistry(Establishedin1903)1926Sumner:Enzymecanbecrystallized1937Krebs:Citricacidcycle(柠檬酸循环)

1944Avery:Pneumonococcustransformationtest1950Pauling:α-helixofprotein2023/2/1821Importantachievement

citricacidcycle(三羧酸循环)

2023/2/1822Importantachievement

citricacidcycleForhisdiscoverythecitricacidcycle

2023/2/1823Historyanddevelopmentancientlybiochemistry(古代生物化学)modernbiochemistry(现代生物化学)1770——1903prepare1903——1953——现在1903Neuberg:Biochemistry(Establishedin1903)1926Sumner:Enzymecanbecrystallized1937Krebs:Citricacidcycle1944Avery:Pneumonococcustransformationtest(肺炎双球菌转化试验)1950Pauling:α-helixofprotein2023/2/1824ImportantachievementTransformation1944Avery,etal.2023/2/1825Historyanddevelopmentancientlybiochemistry(古代生物化学)modernbiochemistry(现代生物化学)1770——1903prepare1903——1953——现在1903Neuberg:Biochemistry(Establishedin1903)1926Sumner:Enzymecanbecrystallized1937Krebs:Citricacidcycle1944Avery:Pneumonococcustransformationtest1950Pauling:α-helixofprotein2023/2/1826Importantachievement

alphahelixstructureofprotein

2023/2/1827HistoryanddevelopmentModernbiochemistryhighlightsfrom19531953Watson&Crick:DNAdoublehelix(双螺旋)1955Sanger:DNAsequence1956Kornberg:DNApolymerase(DNA聚合酶)1958Meselson&Stahl:DNAsemi-conservationreplication(DNA半保留复制)1968Crick:Centraldogma(中心法则)1961Jacob&Monod:Operon(操纵子)1972Berg:RecombinantDNA(重组子DNA)1985Mullis:Polymerasechainreaction(PCR)

(聚合酶链反应)2023/2/1828Importantachievement

DNA

doublehelixmodel

1953

2023/2/18292023/2/1830DNA

doublehelixmodel2023/2/1831HistoryanddevelopmentModernbiochemistryhighlightsfrom19531953Watson&Crick:DNAdoublehelix(双螺旋)1955Sanger:DNAsequence

1956Kornberg:DNApolymerase(DNA聚合酶)1958Meselson&Stahl:DNAsemi-conservationreplication(DNA半保留复制)1968Crick:Centraldogma(中心法则)1961Jacob&Monod:Operon(操纵子)1972Berg:RecombinantDNA(重组子DNA)1985Mullis:Polymerasechainreaction(PCR)

(聚合酶链反应)2023/2/1832PhageX174

5375bp2023/2/1833Importantachievement

PhageX174

5375bp

2023/2/1834HistoryanddevelopmentModernbiochemistryhighlightsfrom19531953Watson&Crick:DNAdoublehelix(双螺旋)1955Sanger:DNAsequence

1956Kornberg:DNApolymerase(DNA聚合酶)1958Meselson&Stahl:DNAsemi-conservationreplication(DNA半保留复制)1968Crick:Centraldogma(中心法则)1961Jacob&Monod:Operon(操纵子)1972Berg:RecombinantDNA(重组子DNA)1985Mullis:Polymerasechainreaction(PCR)

(聚合酶链反应)2023/2/1835Importantachievement

biologicalsynthesisofRNAandDNA

(RNA与DNA生物合成)2023/2/1836HistoryanddevelopmentModernbiochemistryhighlightsfrom19531953Watson&Crick:DNAdoublehelix(双螺旋)1955Sanger:DNAsequence1956Kornberg:DNApolymerase(DNA聚合酶)1958Meselson&Stahl:DNAsemi-conservationreplication(DNA半保留复制)

1968Crick:Centraldogma(中心法则)1961Jacob&Monod:Operon(操纵子)1972Berg:RecombinantDNA(重组子DNA)1985Mullis:Polymerasechainreaction(PCR)

(聚合酶链反应)2023/2/1837Importantachievement

DNAsemi-conservationreplicationDNA半保留复制2023/2/1838HistoryanddevelopmentModernbiochemistryhighlightsfrom19531953Watson&Crick:DNAdoublehelix(双螺旋)1955Sanger:DNAsequence1956Kornberg:DNApolymerase(DNA聚合酶)1958Meselson&Stahl:DNAsemi-conservationreplication(DNA半保留复制)1968Cricketal:Centraldogma(中心法则)1961Jacob&Monod:Operon(操纵子)1972Berg:RecombinantDNA(重组子DNA)1985Mullis:Polymerasechainreaction(PCR)

(聚合酶链反应)2023/2/1839Importantachievement

biologicalsynthesisofRNAandDNA

(RNA与DNA生物合成)2023/2/1840Importantachievement

DNAsemi-conservationreplicationDNA半保留复制2023/2/1841GeneticcodeUUUUUUUUUUUUPhe-Phe-Phe-Phe-

ACACACACACACThr-His-Thr-His-Importantachievement

Geneticcodeandproteinsynthesis2023/2/1842Importantachievement

Geneticcodeandproteinsynthesis遗传密码与蛋白质生物合成2023/2/1843Centraldogma2023/2/1844ImportantachievementRNAdependentDNApolymerase(RNA指导的DNA聚合酶)reversetranscriptase(逆转录酶)2023/2/1845DNAReplication复制RNAReplicationReverseTranscription逆转录ProteinTranslation翻译Transcription转录Centraldogma(中心法则)2023/2/1846HistoryanddevelopmentModernbiochemistryhighlightsfrom19531953Watson&Crick:DNAdoublehelix(双螺旋)1955Sanger:DNAsequence1956Kornberg:DNApolymerase(DNA聚合酶)1958Meselson&Stahl:DNAsemi-conservationreplication(DNA半保留复制)1968Crick:Centraldogma(中心法则)

1961Jacob&Monod:Operon(操纵子)1972Berg:RecombinantDNA(重组子DNA)1985Mullis:Polymerasechainreaction(PCR)

(聚合酶链反应)2023/2/1847lacIPOpromoter-operatorlacrepressorlacZlacYlacAb-galactosidasepermeaseacetylaseLACTOSEGLUCOSE+GALACTOSEb-galactosidasethefunctionofthelactose(lac)operonistoproducetheenzymesrequiredtometabolizelactoseforenergywhenitisrequiredbythecellpromoter(启动子)bindsCAPandRNApolymeraseoperator(操纵基因)bindsthelacrepressor(阻遏物)Importantachievement

ThelactoseoperoninE.coli

乳糖操纵子2023/2/1848lacIPOpromoter-operatorlacrepressorlacIPlacZlacYlacAtherepressortetramerbindstotheoperatorandpreventsRNApolymerasefrombindingtothepromoterlacZlacYlacANOTRANSCRIPTIONRNApolRNApolymeraseisblockedfromthepromoterRegulationofthelactoseoperon-negativecontrol2023/2/1849lacIlacrepressorlacZlacYlacAb-galactosidasepermeaseacetylaseRNApolTRANSCRIPTIONANDTRANSLATIONOCCURinactiverepressorthefunctionofthelactose(lac)operonistoproducetheenzymesrequiredtometabolizelactoseforenergywhenitisrequiredbythecellActivationoflacoperontranscription2023/2/1850Importantachievement

Operon2023/2/1851HistoryanddevelopmentModernbiochemistryhighlightsfrom19531953Watson&Crick:DNAdoublehelix(双螺旋)1955Sanger:DNAsequence1956Kornberg:DNApolymerase(DNA聚合酶)1958Meselson&Stahl:DNAsemi-conservationreplication(DNA半保留复制)1968Crick:Centraldogma(中心法则)1961Jacob&Monod:Operon(操纵子)1972Berg:RecombinantDNA(重组子DNA)

1985Mullis:Polymerasechainreaction(PCR)

(聚合酶链反应)2023/2/1852bacteriaare“transformed”withtherecombinantplasmidcoloniesthatgrowintetracycline,butnotinampicillinareisolatedPstIHumanDNAcutwithPstIPPpBR322ampR,tetRpBR322(humanclone)tetRPPampRtetRpBR322DNAcutwithPstIinactivatingtheampRgenetetRtetRcombineandligateImportantachievement

CloningasegmentofDNAintoaplasmidvector2023/2/18532023/2/1854HistoryanddevelopmentModernbiochemistryhighlightsfrom19531953Watson&Crick:DNAdoublehelix(双螺旋)1955Sanger:DNAsequence1956Kornberg:DNApolymerase(DNA聚合酶)1958Meselson&Stahl:DNAsemi-conservationreplication(DNA半保留复制)1968Crick:Centraldogma(中心法则)1961Jacob&Monod:Operon(操纵子)1972Berg:RecombinantDNA(重组子DNA)1985Mullis:Polymerasechainreaction(PCR)

(聚合酶链反应)2023/2/1855PCR

denature-annealing-extendingcycle

变性-退火-延伸循环

step1.denature2023/2/1856PCR

denature-annealing-extendingcycle

step2.annealingPrimer1Primer22023/2/1857Primer1Primer2BiotinBiotin5’3’5’5’3’5’3’3’TaqDNAPolymerasePCR

denature-annealing-extendingcycle

step3.extending2023/2/1858

靶序列PCRproduct

AfteronecycleBiotinBiotin2023/2/1859No.of No.AmpliconCycles CopiesofTarget1 22 43 84 165 326 6420 1,048,57630 1,073,741,8241cycle=2Amplicon2cycle=4Amplicon3cycle=8Amplicon4cycle=16Amplicon5cycle=32Amplicon6cycle=64Amplicon7cycle=128AmpliconPCRproduct

After30cycles2023/2/1860PCR2023/2/1861Importantachievement

Oncogene

癌基因2023/2/1862Importantachievement

proteinphosphorylation

蛋白质磷酸化2023/2/1863Importantachievement

Gproteins2023/2/18642023/2/1865Importantachievement

nitricoxide(一氧化氮)2023/2/1866Importantachievement

Cellcycle(细胞周期)2023/2/1867Importantachievement

programmedcelldeath(细胞凋亡)程序性细胞死亡2023/2/1868Cont

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