1、基因工程,基因工程學,產物: 重組蛋白質,蛋白質,DNA(基因),mRNA,如胰島素, 干擾素等(93億美元,北美,1997),Cut lactose and x-gal,Inducer: IPTG,lactose,Restriction enzyme,Restriction fragment length polymorphism,Nonautonomous retrotransposonsencode no protein,Alu elements contain the sequence AGCT that is recognized by the restriction enzyme
2、AluI. They are present in up to a million copies in the human genome.They can not transpose by themselves. Instead they take advantage of the retrotransposition machinery of other elements, such as LINEs.,Cut lactose and x-gal,Reporter gene,Fusion protein,Fusion protein,The green fluorescent protein
3、 (GFP) is composed of 238 amino acids (26.9 kDa), originally isolated from the jellyfish Aequorea victoria that fluoresces green when exposed to blue light.,Figure 25.05,1.5 微生物之育種,定點突變,隨機突變:UV等致變劑error prone PCR,GFP derivatives,In 2008, Osamu Shimomura, Marty Chalfie and Roger Tsien (錢永建) won the N
4、obel Prize in chemistry.,重組蛋白質在寄主(host)細胞中的表達,E. coli,染色體,質體,基因,優點產量大操作易缺點no post-translationmodificationinclusion bodynot secrete protein,4,000kb,重組蛋白質在寄主細胞中的表達,E. coligram-positive bacteria缺點B. subtilisnot secrete proteinproteaseslimited vector & promoter“signal” has not found yet,Figure 22.23,DNA
5、 library,基因工程學,昆蟲細胞,染色體,基因,細胞核,病毒,基因治療基因食物,receptor,常見的動物細胞寄主,insect cell-baculovirusmammal-retroviruschinese hamster ovary (CHO) cellsimmortal cell,基因工程學,單株抗體生產Hybridoma桃莉羊無細胞核受精卵 + 體細胞細胞核,質體不穩定,Growth inhibition,使用chemostat為bioreactor其產量將遠大於batch。可是為什麼工業界使用batch的比率遠高於chemostat？ 1.Secondary products 2.Genetic instability 3.Operability and reliability 4.Market economics,
