Recombinant formation of plasmids Site of cleavage Host plasmid cleavage by Restriction Endonucleases ◇ Annealing Point of attatchment and annealing Sticky ends Recombinant Specified Genes Plasmid dna
21 Recombinant formation of plasmids
重组DNA技术 ①获得目的基因; ②与克隆载体连接,形成新的重组DNA分子; ③用重组DNA分子转化受体细胞,并能在受体 细胞中复制和遗传; ④对转化子筛选和鉴定。在具体工作中选择哪 ◇条技术路线; ⑤对获得外源基因的细胞或生物体通过培养, 获得所需的遗传性状或表达出所需要的产物。 22
22 重组DNA技术 • ①获得目的基因； • ②与克隆载体连接，形成新的重组DNA分子； • ③用重组DNA分子转化受体细胞，并能在受体 细胞中复制和遗传； • ④对转化子筛选和鉴定。在具体工作中选择哪 条技术路线； • ⑤对获得外源基因的细胞或生物体通过培养， 获得所需的遗传性状或表达出所需要的产物
Synthetic insulin production using recombinant dNa ◇· One of the biggest breakthroughs in recombinant dNa technology happened in the manufacture of biosynthetic human insulin which was the first medicine made ◇ via recombinant dna technology ever to be approved by the FDa in 1982 23
23 Synthetic insulin production using recombinant DNA • One of the biggest breakthroughs in recombinant DNA technology happened in the manufacture of biosynthetic "human" insulin, which was the first medicine made via recombinant DNA technology ever to be approved by the FDA in 1982
Human recombinants with recombination as only source ◆· Erythropoietin(EPO促红细胞生成素) 2 Epogen from ◇ Amgen Granulocyte colony- stimulating factor(G-CSF粒细胞 集落刺激因子) filgrastim sold as Neupogen from Amgen; pegfilgrastim sold as neulasta Fabrazyme by genzyme Alpha-L- iduronidase( rhIDU; laronidase艾杜糖苷酸酶) ◇ Aldurazyme by Biomarin Pharmaceutical and Genzyme N-acetylgalactosamine-4-sulfatase ( rhASB; galsulfaseN-乙酰半乳糖胺硫酸酯酶) Naglazyme(TM by biomarin pharmaceutical 24
24 Human recombinants with recombination as only source • Erythropoietin (EPO促红细胞生成素) Epogen from Amgen • Granulocyte colony-stimulating factor (G-CSF粒细胞 集落刺激因子) filgrastim sold as Neupogen from Amgen; pegfilgrastim sold as Neulasta • Fabrazyme by Genzyme • Alpha-L-iduronidase (rhIDU; laronidase艾杜糖苷酸酶) Aldurazyme by Biomarin Pharmaceutical and Genzyme • N-acetylgalactosamine-4-sulfatase (rhASB; galsulfaseN-乙酰半乳糖胺硫酸酯酶) Naglazyme (TM) by Biomarin Pharmaceutical
Human recombinants with recombination as only source Dornase alfa(阿法链道酶<囊性纤维变性病治疗芶 ) a dnase sold under the trade name pulmozyme by genentech issue plaminogen activator(TPA) Activase by Genentech Glucoceret; rosidae ceredase(葡萄糖脑苷脂酶)by Genzyme ◇ Interferon(If Interferon-beta-la as avonex from Biogen Idec: Rebif from Serono Interferon beta-l b as Betaseron from Schering nsulin-like growth factor 1(IGF-1
25 • Dornase alfa（阿法链道酶<囊性纤维变性病治疗药 >）, a DNase sold under the trade name Pulmozyme by Genentech • Tissue plaminogen activator (TPA) Activase by Genentech • Glucocerebrosidase Ceredase（葡萄糖脑苷脂酶） by Genzyme • Interferon (IF) Interferon-beta-1a as Avonex from Biogen Idec; Rebif from Serono;Interferon beta-1b as Betaseron from Schering • Insulin-like growth factor 1 (IGF-1) Human recombinants with recombination as only source