PCR primer design ATTGCCACGTTGACGGTCAG CACGATTTCCGGAAGTAC Inducer IPTG lactose
- Slides: 42
PCR: primer design ATTGCCACGTTGACGGTCAG CACGATTTCCGGAAGTAC
Inducer: IPTG, lactose
Restriction endonucleases • Palindromes • Blunt ends v. s. sticky ends
重組蛋白質 在寄主(host)細胞中的表達 基因 優點 • 產量大 • 操作易 質體 染色體 4, 000 kb E. coli 缺點 • no post-translation modification • inclusion body • not secrete protein
重組蛋白質 在寄主細胞中的表達 E. coli 缺點 not secrete protein gram-positive bacteria B. subtilis proteases limited vector & promoter “signal” has not found yet
Growth inhibition
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 Infection Gene Expression and Protein Synthesis: • Immediately-Early Genes • Delay-Early Genes • Late Genes • Very Late Genes: polyhedrin and p 10 Virus Assembly and Release
Infect cultured insect cells Cotransfection with Ac. MNPV DNA Amplify and titer the recombinant virus Confirm presence of target gene by PCR or Southern blot Plaque Assay for putative recombinant virus
Generation of Recombinant Baculovirus by Bacmid
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 R+ A- + B - R+ B - + A -
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
The Sanger dideoxy method of DNA sequencing Figure 5. 18
Molecular tools • • Gel Electrophoresis Labeled Tracers Hybridization Mapping and quantifying transcripts Measuring transcription rates in vivo Assaying DNA-protein interactions Knockouts
S 1 mapping the 5’-end of a transcript
Primer extension
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
Figure 5. 35
DNase footprinting Page 125
Molecular tools • • Gel Electrophoresis Labeled Tracers Hybridization Mapping and quantifying transcripts Measuring transcription rates in vivo Assaying DNA-protein interactions Knockouts
- Sparqed
- Fret rt pcr
- Drug metabolism definition
- Glucouronide
- Hme inducer
- Enzyme inducer drugs
- Cheese lactose chart
- Pont oxydique
- Lactose structure
- Causes of lactose intolerance
- Hemiacetal group in lactose
- Lactose intolerance by ethnicity
- P mirabilis lactose fermentation
- Lactose acetal and hemiacetal
- Reducing properties of monosaccharides
- Lactose is a disaccharide found in milk
- Lactose
- Lactose positiva
- Lactosen
- Lactose intolerant map
- Lactose
- Lactose glycemic index
- Biologi
- Lactobacillus bulgaricus converts lactose to *
- Proteus non lactose fermenter
- Lupinesse
- Fisiopatologia intolerancia a lactose
- Double strength lactose broth
- Lactose intolerance histology
- Fructose is a ketohexose
- Lactose free milk
- Structure of sucrose maltose and lactose
- Lactose intolerance gene
- Lactose test strips
- Enterobacteriaceae non lactose fermenter
- Symptoms of lactose intolerance
- Enfamil ar 24 calorie formula recipe
- Colony pcr yeast
- Fases pcr
- Retrotranscripción
- Rcp extracorpórea
- Poonum patel
- Pcr phases