A plasmid vector for gene expression Expression vectors: allowing the exogenous dna to be inserted stored and expressed 1 Promoter and terminator for rNa transcription are required 2. Intact ORF and ribosomal binding sites (RBS)are required for translation 3. Include:(1)bacterial expression vectors, (2) yeast expression vectors, (3)mammalian expression vector
A plasmid vector for gene expression Expression vectors: allowing the exogenous DNA to be inserted, stored and expressed. 1. Promoter and terminator for RNA transcription are required. 2. Intact ORF and ribosomal binding sites (RBS) are required for translation. 3. Include：(1) bacterial expression vectors, (2) yeast expression vectors, (3) mammalian expression vector
T7 promoter s RBS Start codon MCS Ampr Transcription terminator expression vector or An bacterial expression vector
T7 promoter RBS Start codon MCS Transcription terminator Ampr ori T7 expression vector An bacterial expression vector
MCS ATG 6xHis Xpress"Epitope EK Site V5 epitope 6xHis term C C YES Vectors 3 TRP A yeast expression vector
MCS A yeast expression vector
Bacteriophage vector Two examples: 1.A phage bacteriophage 人 replacement vector 2. M13 phage M13 phage vector Cloning in M13 Hybrid plasmid -M13 vectors
Bacteriophage vector Two examples: 1.λ phage • bacteriophageλ • λ replacement vector 2.M13 phage • M13 phage vector • Cloning in M13 • Hybrid plasmid-M13 vectors
A-Phage genome Repl aceable region A phage DNA recombination viruses that can Target DNA nfect bacteria Packaging .48. 5 kb in length assembly Linear or circular genome(cos ends) Lytic phase (Replicate and Infect E. coli release Lysogenic phase (integrate into host genome
• viruses that can infect bacteria. •48.5 kb in length •Linear or circular genome (cos ends) •Lytic phase (Replicate and release) •Lysogenic phase (integrate into host genome) λ phage