IntroductiontoBiologicalScience生物科技概论ChungW

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1、1 Introduction toBiological Science生物科技概論By:Chung W.Wu,M.S.,R.Ph Ext:-52132Ancient Biological Science1.Food Preparation Beer,Cheese2.Fermentation Microbial Food-Perfume3.Alcohol Fermentation Distillation Alcohol Concentration31st World War Germany Glucose-Glycerol British Sugar-Acetone+Butanol Clost

2、ridium acetobutylicum Carbon Dioxide Hydrogen Gas:Explosive 42nd World War1940 FlemingPenicillium notatum Penicillin:Broad Spectrum Antibiotic 2mg/lPenicillium chrysogenum UV,X-RayMutation 20mg/lStreptomyces griseus Stretomycin5Genetic EngineeringDNA Recombination RecombinantDeoxyribonucleic AcidChe

3、mical Composition1.Phosphate2.Pentose3.Nitrogenous Base Adenine Thymine Cytosine Guanine6Principles of DNA RecombinantDNA Section cant Replicate by itself.1.Cloning Vector 具有複製起始能力之載體2.Plasmid 能獨立複製之環狀DNA分子3.Restriction Endonuclease (DNase)Recognition Site4.Sticky Ends7Expression of Cloned Genes1.Tr

4、anscription2.Translation3.Post-Translation 8The Molecular Basis of Heredity:DNA&RNANuclear ComponentsDNATranscriptionRNATranslationAmino AcidsPolypeptideProteins RNA:Single Strand Adenine,Uracil,Cytosine,Guanine mRNA t RNA r RNA9 DNA Transcription1.Duplex DNA Seperation 2 Single DNA Strands Exposing

5、 the Nitrogenous Base Sequences Only One of the 2 DNA Strands,Which Serves as A Template,or Pattern(Gene),for the Synthesis of RNA.10 2.ComplementaryTTT TCT AAACGTAAAAGATTT GCAmRNAUUUUCUAAACGU4x4x4=6464 Possible 3-Letter Words11TranslationProtein Synthesis20 Words in the Formof 20 Common A.A.Each Tr

6、iplet Codes foran Amino Acids12Ribosome:Synthesis of ProteinCellular Organelle thatServes as the Meeting Place for mRNA andtRNA that carry A.A.to Form PeptideUUUUCUAAACGUPhe-Ser-LysArg13What is Gene Cloning?Insert DNA Fragment toCircle DNA Molecule-Chimera Recombinant DNA14PCR:Polymerase Chain React

7、ion1995 Kary Mullis1.Denaturation of DNA 94C2.Annealing of the Primers(Oligonucleotides)50-60C 3.Synthesis of New DNA 74C4.Renaturation of DNARepeat the Cycle 25-30 Times15Gene Isolation by Cloning1.DNA Fragments2.Construct Recombinant DNA3.Introduce into Bacteria4.Plate Out5.Each Colony contains Mu

8、ltiple Copies of just one 16Gene Isolation by PCR1.Polymerase Chain Reaction2.X Several Million17The Commercial Exploitation of Microorganism 1.Bacillus thuringiensis Natural Insecticide Thuringiensis Gene 選殖共生於玉米根部 可以防治玉米害蟲182.Aureobasidium pullulans Polysaccharides Pullulan193.Acinetobacter calcoa

9、ccticus Polysaccharides Emulsan20Biotransformation Bioconversion1.Steroid-Cortisone 37 steps USD 200 dollars/g 11 steps USD 6 dollars/g21 GlucoseSorbitol-Sorbose -Vitamin C GlucoseSorbitol 4 chemical steps Biotransformation 1 chemical step22Examples:1.Phenylpyruvic Acid -Phenylalanine 2.Cinnamic aci

10、d-Phenylalanine Phe+Aspartic Acid-Aspartame 160 X Sweetness23 BiotransformationGrowing Cell休眠Cell or Killed Cell24 Enzymes:1.Glucose-Fructose Glucose Isomerase Arthrobacter sp.Bacillus sp.252.Penicillin-6-APC Penicillin Amidase Escherichia coli Through DNA Recombinant,the yield Of Enzyme:20g/l263.Ch

11、ymosin 小牛胃中凝乳素 Milk-Cheese (with Flavor)27Protein:1.Growth Hormone Fish Cow:secretion of milk Human282.Insulin3.Interferon4.VaccineGMP Requirement29Cell and Enzyme ImmobilizationMethods of Immobilization1.Physical Adsorption Physical Binding Cell or Enzyme以吸附方式被固定裝置在 管柱中容易吸附也容易脫離尤其在基質添加時Cell or Enzy

12、me容易Autolyse造成Activity 喪失30 2.Cross linking 以Glutaraldehyde Toluene diisocyanate 進行交聯形成不溶解性間質但Cell or Enzyme Activity仍會流失313.Entrapment Lattice Collagen Gelatin,Agar Polyacrylamide CelluloseTo form Polymer Matrix 32Choice of Immobilization Methods Stability Cost Regenerability Commercialization33Mon

13、oclonal Antibody Polyclonal Antibody Epitopes 抗原決定位 Chimeric 嵌人化 Humanized 擬人化34Total Human Monoclonal AntibodyMaking A HybridomaAntigen-AnimalImmunizationBlood Collection Speen of Animal RBC&Lymphocyte35Application of Monoclonal Antibody1.Diagnostic Kit2.增加敏感度與再現性3.Therapy36Examples:1.Hepatitis Vir

14、us2.Polyoma3.Adenovirus 37Isolation&Purification of MABsImmunoaffinity Chromatography1.Specificity2.Elution-Sharp Peak3.Independent Target Protein38Immunotoxins as Therapeutic Ricin-A,B Polypeptide -S-S-A,B Chain Enter Cell PlasmRibosome-Dysfunction-Cells Death39Immunotoxin:MABs combine with A,B Chain -Attack Cancer Cell Stable in Human Body402.MABs+Radioisotope 目標導向 放射線能殺死癌細胞 Radioisotope:High Energy Low Penetration Short Half Life Safety Ex:At 13hrs Y 64hrs41Human Gene TherapySomatic Cell Gene TherapyCorrection of Germ Cell 1.Hard To Isolation 2.Hurt Normal Cell