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【正文】 丙氨膦（ Bialaphos） 204， 土霉素 205 反向遺傳學(xué)（ Reverse geics） 利福霉素 206， 轉(zhuǎn)座子誘變（ Transposon mutagenesis） 達(dá)托霉素（ Daptomycin） 37 蛋白組學(xué)（ Proteomics） 尼柯霉素（ Nikkomycin） 114 異源探針（ Heterologous probes） II型 PKS130， I型 PKS154， 脫氧糖 141 PCR和同源探針 II型 PKS139， 脫氧糖 137， 138， 207， 208， NRPS 140， 3氨基 5羥苯甲酸合酶 137， 鹵化酶（ halogenase） 209， III型 PKS 134 抗生素生物合成基因克隆策略 1. 將目的基因克隆到標(biāo)準(zhǔn)宿主菌，然后檢測個別基因產(chǎn)物。 鏈霉菌宿主菌 ? Streptomyces lividans TK132TK2 TK64等 ? Streptomyces coelicolor A3(2) ? Other Streptomyces Vectors for Streptomyces ? 高拷貝載體 :pIJ101, pIJ702 ? 低拷貝載體 :pIJ61, pIJ922, pIJ941 ? 穿梭載體 : pIJ699 ? Cosmid:pKC505 ? 接合轉(zhuǎn)移載體 :pPM801 ? 噬菌體載體 :fC31系列 ? 表達(dá)載體 :pIJ6021, pIJ4123 ? 分泌載體 Other vectors ? 大容量載體：采用酵母人工染色體（ YAC）克隆技術(shù)構(gòu)建了細(xì)菌 BAC載體，插入片段 100~300kb。m wide, typical for bacteria and much smaller than fungal hyphae and spores. (Scanning electron micrograph, Mark Buttner, Kim Findlay, John Innes Centre). Close up of Streptomyces coelicolor spores Close up of Streptomyces coelicolor spores (c. 1181。 the reddish mutant colonies are not forming aerial mycelium. The red mycelium colour and the dark background is from the antibiotics produced by Streptomyces coelicolor (John Innes Centre, Photography Department). Single colony of Streptomyces coelicolor Single colony of Streptomyces coelicolor topped by a liquid droplet containing antbiotic. Liquid droplets are often found in Streptomyces colonies, it is not clear how they are generated. (John Innes Centre, Photography Department). Close up of a colony of Streptomyces coelicolor mutant D132 Close up of a colony of Streptomyces coelicolor mutant D132 which is produces undecylprodigiosin but not actinorhodin (image provided by Karen Jolly, University of Leeds). Mass of curled aerial mycelium of Streptomyces coelicolor Mass of curled aerial mycelium (c. 1 181。m wide) of Streptomyces grown in liquid broth (light microscope 。工業(yè)微生物菌種選育技術(shù) 鏈霉菌基因工程 為什么鏈霉菌？ 鏈霉菌的重要性 ? 產(chǎn)生絕大多數(shù)已知的抗生素 ? 生物合成機制的闡明 ? 生產(chǎn)工業(yè)用酶的潛力 – 葡萄糖異構(gòu)酶 – 蛋白酶 – 纖維素酶 – 木聚糖酶 – 。 What Are Streptomyces? ? They are Gram Positive filamentous Bacteria. ? They are obligate aerobic bacteria that are abundant in most soils. ? Streptomyces have two metabolic stages, Primary and Secondary Metabolism. ? They utilize insoluble anic debris by producing a variety of extracellular hydrolytic enzymes. 放線菌生活史 Streptomyces coelicolor colonies with aerial mycelium and spores The area of the picture represents about 2x3 cm. The blue haloes around the colonies are secreted a c t i n o r h o d i n . A c t i n o r h o d i n is an a n t i b i o t i c ( n o t u s e d clinically) which is blue under alkaline conditions and red under acidic conditions. Actinorhodin is a polyketide made by multiple condensations of acetate by a Type II p o l y k e t i d e s y n t h a s e (John Innes Centre, Photography Department). Young vegetative mycelium Youn
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