{"title":"双歧链霉菌染色体和膜部分共价闭合环状DNA的分离与鉴定。","authors":"H Ikeda, H Tanaka, S Omura","doi":"10.7164/antibiotics.35.497","DOIUrl":null,"url":null,"abstract":"<p><p>Covalently closed circular (ccc) DNAs were isolated by a technique involving alkaline denaturation from the spiramycin producer Streptomyces ambofaciens KA-1028 and also from spiramycin non-producing strains AF-11 and QN-25; plasmids could not be detected in these strains by a cleared lysate method. It was found that most of the ccc DNA in these strains was present in the chromosomal and membrane fractions. These ccc DNAs had identical mobilities in agarose gel electrophoresis. The size was calculated to be 53.1 x 10(6) daltons from the contour length measurements. The ccc DNA gave one fragment on digestion with Hind III, three fragments with Eco R1, and twenty-eight fragments with Bam H1.</p>","PeriodicalId":501839,"journal":{"name":"The Journal of Antibiotics","volume":" ","pages":"497-506"},"PeriodicalIF":0.0000,"publicationDate":"1982-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.7164/antibiotics.35.497","citationCount":"12","resultStr":"{\"title\":\"Isolation and characterization of covalently closed circular DNA associated with chromosomal and membrane fraction from Streptomyces ambofaciens.\",\"authors\":\"H Ikeda, H Tanaka, S Omura\",\"doi\":\"10.7164/antibiotics.35.497\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Covalently closed circular (ccc) DNAs were isolated by a technique involving alkaline denaturation from the spiramycin producer Streptomyces ambofaciens KA-1028 and also from spiramycin non-producing strains AF-11 and QN-25; plasmids could not be detected in these strains by a cleared lysate method. It was found that most of the ccc DNA in these strains was present in the chromosomal and membrane fractions. These ccc DNAs had identical mobilities in agarose gel electrophoresis. The size was calculated to be 53.1 x 10(6) daltons from the contour length measurements. The ccc DNA gave one fragment on digestion with Hind III, three fragments with Eco R1, and twenty-eight fragments with Bam H1.</p>\",\"PeriodicalId\":501839,\"journal\":{\"name\":\"The Journal of Antibiotics\",\"volume\":\" \",\"pages\":\"497-506\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"1982-04-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.7164/antibiotics.35.497\",\"citationCount\":\"12\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"The Journal of Antibiotics\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.7164/antibiotics.35.497\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"The Journal of Antibiotics","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.7164/antibiotics.35.497","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 12
摘要
用碱性变性技术从产螺旋霉素的双歧杆菌链霉菌KA-1028和不产螺旋霉素的菌株AF-11和QN-25中分离出共价闭合环状(ccc) dna;在这些菌株中,通过清除裂解法无法检测到质粒。发现这些菌株的大部分ccc DNA存在于染色体和膜部分。这些ccc dna在琼脂糖凝胶电泳中具有相同的迁移率。根据轮廓长度测量计算出尺寸为53.1 x 10(6)道尔顿。Hind III酶切得到1个片段,Eco R1酶切得到3个片段,Bam H1酶切得到28个片段。
Isolation and characterization of covalently closed circular DNA associated with chromosomal and membrane fraction from Streptomyces ambofaciens.
Covalently closed circular (ccc) DNAs were isolated by a technique involving alkaline denaturation from the spiramycin producer Streptomyces ambofaciens KA-1028 and also from spiramycin non-producing strains AF-11 and QN-25; plasmids could not be detected in these strains by a cleared lysate method. It was found that most of the ccc DNA in these strains was present in the chromosomal and membrane fractions. These ccc DNAs had identical mobilities in agarose gel electrophoresis. The size was calculated to be 53.1 x 10(6) daltons from the contour length measurements. The ccc DNA gave one fragment on digestion with Hind III, three fragments with Eco R1, and twenty-eight fragments with Bam H1.