基因工程學.ppt

基因工程學 Restriction endonucleases Palindromes Blunt ends v.s. sticky ends 重組蛋白質在寄主(host)細胞中的表達 重組蛋白質在寄主細胞中的表達 E. coli gram-positive bacteria 缺點 B. subtilis not secrete protein proteases limited vector promoter “signal” has not found yet Growth inhibition 基因工程學 常見的動物細胞寄主 insect cell-baculovirus mammal-retrovirus chinese hamster ovary (CHO) cells immortal cell Baculovirus Replication and Gene Expression Molecular tools Gel Electrophoresis Labeled Tracers Hybridization Mapping and quantifying transcripts Measuring transcription rates in vivo Assaying DNA-protein interactions Knockouts Pulse-field gel electrophoresis(0.2-2.2 Mb) Two-dimensional gel electrophoresis Ion exchange chromatography Gel filtration chromatography Affinity chromatography Molecular tools Gel Electrophoresis Labeled Tracers Hybridization Mapping and quantifying transcripts Measuring transcription rates in vivo Assaying DNA-protein interactions Knockouts Molecular tools Gel Electrophoresis Labeled Tracers Hybridization Mapping and quantifying transcripts Measuring transcription rates in vivo Assaying DNA-protein interactions Knockouts DNA fingerprinting Molecular tools Gel Electrophoresis Labeled Tracers Hybridization Mapping and quantifying transcripts Measuring transcription rates in vivo Assaying DNA-protein interactions Knockouts Molecular tools Gel Electrophoresis Labeled Tracers Hybridization Mapping and quantifying transcripts Measuring transcription rates in vivo Assaying DNA-protein interactions Knockouts Molecular tools Gel Electrophoresis Labeled Tracers Hybridization Mapping and quantifying transcripts Measuring transcription rates in vivo Assaying DNA-protein interactions Knockouts Molecular tools Gel Electrophoresis Labeled Tracers Hybridization Mapping and quantifying transcripts Measuring transcription rates in vivo Assaying DNA-protein interactions Knockouts S1 mapping the 5’-end of a transcript Primer extension Figure 5.35 DNase footprinting Page 125 * 精品P