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基因在大肠杆菌、酵母中的高效表达(High expression of genes in Escherichia coli and yeast).doc

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基因在大肠杆菌、酵母中的高效表达(High expression of genes in Escherichia coli and yeast) [teaching catalogue and class assignment] Expression system and expression strategy of 4.1 gene (0.5) High expression of 4.2 gene in Escherichia coli (2) 4.2.1 Escherichia coli expression system based on T7 phage RNA polymerase / promoter Fusion expression of 4.2.2 protein Secreted expression of 4.2.3 protein Inclusion bodies, expression and protein refolding of 4.2.4 proteins High expression of 4.3 gene in yeast (1.5) 4.3.1 yeast expression system overview 4.3.2 methanol yeast expression system Expression of 4.3.3 tissue plasminogen activator in Pichia pastoris [teaching purposes] Enable students to master the concepts of gene expression systems, efficient expression strategies for genes in Escherichia coli and yeasts, and examples of gene efficient expression. [emphasis on teaching] Strategies and methods for efficient expression of genes in Escherichia coli. [teaching difficulties] High expression of genes in yeast. [teaching mode] Multimedia interpretation combined with elicitation and discussion teaching [teaching process and content] Section 1 gene expression system and expression strategy (0.5) Gene expression system Gene expression is the product of a purposeful synthesis of the target gene. Including transcription, translation, processing, and other processes. Expression vector: the target gene is placed in a biological host under artificial control and produced in large quantities. Three systems of expressing vectors: 1, DNA replication and plasmid DNA screening: there are multiple cloning sites, DNA replication starting point ori, and Amp, Tet resistance genes 2 regulation of transcription of target genes: this system includes promoters, inhibitors, genes, and transcription terminators. Promoters are located upstream of the target gene and are commonly used, such as Placz 3 protein translation, including ribosome recognition site SD, translation initiation codon and terminator codon
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