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首页> 外文期刊>The Journal of Antibiotics: An International Journal >Cloning and heterologous expression of genes from the kinamycin biosynthetic pathway of Streptomyces murayamaensis
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Cloning and heterologous expression of genes from the kinamycin biosynthetic pathway of Streptomyces murayamaensis

机译：村山链霉菌金霉素生物合成途径基因的克隆和异源表达

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摘要

The genes for most of the biosynthesis of the kinamycin antibiotics have been cloned and heterologously expressed. Genomic DNA of Streptomyces murayamaensis was partially digested with MboI and a library of similar to 40 kb fragments in E. coli XL1-BlueMR was prepared using the cosmid vector pOJ446. Hybridization with the actI probe from the actinorhodin polyketide synthase genes identified two clusters of polyketide genes. After transferal of these clusters to S. lividans ZX7, expression of one cluster was established by HPLC with photodiode array detection. Peaks were identified from the kin cluster for dehydrorabelomycin, kinobscurinone, and stealthin C, which are known intermediates in kinamycin biosynthesis. Two shunt metabolites, kinafluorenone and seongomycin were also identified. The structure of the latter was determined from a quantity obtained from large-scale fermentation of one of the clones. [References: 39]
机译：已克隆并异源表达了大多数卡那霉素抗生素的生物合成基因。用MboI部分消化村山链霉菌的基因组DNA，并使用粘粒载体pOJ446制备了大肠杆菌XL1-BlueMR中约40 kb片段的文库。与来自放线菌素聚酮化合物合酶基因的actI探针的杂交鉴定了两个聚酮化合物基因簇。将这些簇转移至紫葡萄链球菌ZX7后，通过具有光电二极管阵列检测的HPLC建立一个簇的表达。从亲缘簇中确定了脱氢雷贝霉素，基诺布斯酮和隐身蛋白C的峰，这是在卡那霉素生物合成中已知的中间体。还鉴定了两种分流代谢物，基那芴酮和Seongomycin。后者的结构由克隆之一的大规模发酵获得的量确定。 [参考：39]

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