Targeting polyketide synthase gene pool within actinomycetes: new degenerate primers

Savic M; Vasiljevic B

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Targeting polyketide synthase gene pool within actinomycetes: new degenerate primers

机译：放线菌内靶向聚酮化合物合酶基因库：新的简并引物

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Natural products provide a unique element of molecular diversity and biological functionality and they are still indispensable for drug discovery. The polyketides, comprising a large and structurally diverse family of bioactive natural products, have been isolated from a group of mycelia-forming Gram-positive microorganisms, the actinomycetes. Relatively high amino acid sequence identity of the actinomycetes type I polyketide synthases (PKSs) was used to design three degenerate primer pairs for homology-based PCR detection of novel PKS genes, with particular interest into PKSs involved in biosynthesis of immunosuppressive-like metabolites. The stepdown PCR method, described here, enables fast insight into the PKS arsenal within actinomycetes. Designed primers and stepdown PCR were applied for the analysis of two natural isolates, Streptomyces sp. strains NP13 and MS405. Sequence analysis of chosen clones revealed the presence of two distinctive sequences in strain Streptomyces sp. NP13, but only one of these showed homology to PKS-related sequences. On analysing PCR amplicons derived from Streptomyces sp. strain MS405, three different PKS-related sequences were identified demonstrating a potential of designed primers to target PKS gene pool within single organism.

机译：天然产物提供了分子多样性和生物学功能的独特元素，它们仍然是药物发现必不可少的。从一组形成菌丝体的革兰氏阳性微生物，放线菌中分离出包含大分子和结构多样的生物活性天然产物家族的聚酮化合物。放线菌I型聚酮化合物合酶（PKSs）的氨基酸序列相对较高，被用于设计三个简并引物对，用于基于同源性的PCR检测新的PKS基因，特别是涉及参与免疫抑制样代谢物生物合成的PKS中。本文所述的逐步PCR方法可快速了解放线菌内的PKS武库。将设计的引物和逐步PCR用于两种自然分离株链霉菌的分析。菌株NP13和MS405。所选克隆的序列分析揭示了在链霉菌属菌株中存在两个独特的序列。 NP13，但只有其中之一显示出与PKS相关序列的同源性。在分析源自链霉菌属的PCR扩增子。确定了MS405菌株MS405的三个不同的PKS相关序列，这表明设计的引物有可能靶向单一生物体内的PKS基因库。

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《Journal of industrial microbiology & biotechnology》 |2006年第6期|共8页
作者
Savic M; Vasiljevic B;
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正文语种 eng
中图分类工业微生物学;
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Actinobacteria enzymology genetics; Amino Acid Sequence; Blotting; Southern; Chromatography; Thin Layer; DNA Primers; DNA; Bacterial analysis chemistry genetics; Genes; Bacterial; Macrolides analysis isolation amp; purification; Molecular Sequence Data; Phylogeny; Polyketide Synthases genetics; Polymerase Chain Reaction methods; Research Support; Non-U.S. Gov't; Sequence Analysis; DNA; Sequence Homology; Amino Acid;

机译：放线菌的酶遗传学;氨基酸序列;印迹;Southern;色谱;薄层;DNA引物;DNA;细菌分析化学遗传学;基因;细菌;杀虫剂分析的分离和纯化;分子序列数据;系统发育;聚酮合酶遗传学;聚合酶链反应方法;研究支持;非美国政府;序列分析;DNA;序列同源性;氨基酸;

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1. Targeting polyketide synthase gene pool within actinomycetes: new degenerate primers [J] . Savic M, Vasiljevic B Journal of industrial microbiology & biotechnology . 2006,第6期

机译：放线菌内靶向聚酮化合物合酶基因库：新的简并引物
2. A Sorangium cellulosum (myxobacterium) gene cluster for the biosynthesis of the macrolide antibiotic soraphen A: cloning, characterization, and homology to polyketide synthase genes from actinomycetes. [J] . T Schupp, C Toupet, B Cluzel, Journal of bacteriology . 1995,第13期

机译：用于大环内酯类抗生素soraphen A的生物合成的纤维素膜（粘细菌）基因簇：克隆，表征和与放线菌中聚酮化合物合酶基因的同源性。
3. A Sorangium cellulosum (myxobacterium) gene cluster for the biosynthesis of the macrolide antibiotic soraphen A: cloning, characterization, and homology to polyketide synthase genes from actinomycetes. [J] . T Schupp, C Toupet, B Cluzel, Journal of bacteriology . 1995,第13期

机译：用于大环内酯类抗生素soraphen A的生物合成的纤维素膜（粘细菌）基因簇：克隆，表征和与放线菌中聚酮化合物合酶基因的同源性。
4. Development of Real-Time PCR Primer and Probe Sets for Detecting Degenerated and Non-degenerated Forms of the Butanol-Producing Bacterium Clostridium acetobutylicum ATCC 824 [C] . Sun-Mi Lee, Min Ok Cho, Youngsoon Um, Symposium on biotechnology for fuels and chemicals . 2010

机译：用于检测丁醇产物脱苯乙烯酸梭菌ATCC 824的丁醇产物菌梭菌梭菌的退化和非退化形式的实时PCR引物和探针组
5. En masse cloning of expressed disease resistance genes of wheat (Triticum aestivum) using RNA differential display via degenerate primers and data mining methods [D] . Dilbirligi, Muharrem 2004

机译：通过简并引物和数据挖掘方法利用RNA差异显示技术大规模克隆小麦（Triticum aestivum）表达的抗病基因
6. PCR effects of melting temperature adjustment of individual primers in degenerate primer pools [O] . Ankur Naqib, Trisha Jeon, Kevin Kunstman, -1

机译：简并引物库中单个引物的解链温度调节的PCR效应
7. A Sorangium cellulosum (myxobacterium) gene cluster for the biosynthesis of the macrolide antibiotic soraphen A: cloning, characterization, and homology to polyketide synthase genes from actinomycetes. [O] . Schupp, T, Toupet, C, Cluzel, B, 1995

机译：用于大环内酯类抗生素soraphen A的生物合成的纤维素膜（粘杆菌）基因簇：克隆，表征和与放线菌中聚酮化合物合酶基因的同源性。

Targeting polyketide synthase gene pool within actinomycetes: new degenerate primers

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