基因工程
基因工程學 產物: 重組蛋白質 DNA(基因) mRNA 蛋白質 如胰島素, 干擾素等(93億美元,北美,1997) 胰島素缺乏 糖尿病
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Cut lactose and x-gal
Inducer: IPTG, lactose
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Restriction enzyme 8
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Restriction fragment length polymorphism 13
Nonautonomous retrotransposons encode no protein Alu elements contain the sequence AGCT that is recognized by the restriction enzyme 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.
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Cut lactose and x-gal
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Reporter gene 20
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Fusion protein
Fusion protein The green fluorescent protein (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.
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Figure 25.05 25
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 Nobel Prize in chemistry.
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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 缺點 no post-translation modification inclusion body not secrete protein 質體 染色體 4,000kb E. coli
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
DNA library Figure 22.23 33
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基因工程學 基因 病毒 receptor 染色體 細胞核 基因治療 基因食物 昆蟲細胞
常見的動物細胞寄主 insect cell-baculovirus mammal-retrovirus chinese hamster ovary (CHO) cells immortal 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
3.Operability and reliability 4.Market economics 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