期刊文献+

红色糖多孢菌ΔSACE_0065,ΔSACE_2559和ΔSACE_2565突变体构建及对孢子分化的影响

ΔSACE_0065,ΔSACE_2559 and ΔSACE_2565 genes mutation in Saccharopolyspora erythraea and their effect on spore differentiation
原文传递
导出
摘要 目的构建红色糖多孢菌ΔSACE_0065、ΔSACE_2559和ΔSACE_2565三个突变体并研究其功能。方法将待失活目的基因的上下游同源片段依次连接到pUCTSR质粒Thio抗性基因(tsr)两侧,利用PEG介导的原生质体转化法将线性DNA同源片段转入红色糖多孢菌A226内,通过同源重组使目的基因失活,最后将构建出的三个突变株与出发菌株A226及ΔbldD突变株进行比较,观察孢子生长有无差异。结果与结论成功构建ΔSACE_0065、ΔSACE_2559和ΔSACE_2565三个突变株;与A226相比,ΔSACE_0065和ΔSACE_2559气生菌丝和孢子形成推迟,而ΔSACE_2565突变株的生长情况则无明显变化,提示红色糖多孢菌SACE_0065、SACE_2559基因参与其形态分化的调控。 Objective To construct the mutant of Saccharopolyspora erythraea in which SACE_0065,SACE_2559 or SACE_2565 gene is knocked out respectively,and to make preliminary function analysis of those genes.Methods Linear DNA containing thiostrepton resistance gene(tsr) with upper and down homologous fragments of the target inactivated gene around tsr was transformed into Sac.erythraea A226 under PEG mediation.By chromosomic homologous recombination,the mutant in which the target gene was disrupted could be screened out with thiostrepton choice.The mutant strains were compared with the original strain A226 and the ΔbldD mutant in differentiation of aerial hyphae into spores.Results and Conclusion The growth of aerial hyphae and the differentiation of spores of ΔSACE_0065 and ΔSACE_2559 mutants were delayed,while ΔSACE_2565 mutant did not change obviously compared with the original strain A226.SACE_0065 and SACE_2559 in Sac.erythraea may be involved in regulating the morphological differentiation,and could be regulatory genes in the development of spores.
出处 《军事医学》 CAS CSCD 北大核心 2011年第12期913-916,共4页 Military Medical Sciences
基金 国家大学生创新性实验计划项目(091035711) 国家自然科学基金资助项目(30870069)
关键词 红色糖多孢菌 SACE_0065 SACE_2559 SACE_2565 bldD 孢子分化 Saccharopolyspora erythraea SACE_0065 SACE_2559 SACE_2565 bldD spore differentiation
  • 相关文献

参考文献10

  • 1刘惠,黄训端,刘道琴,赵玮,樊伟,韩姝,张部昌.红色糖多孢菌染色体基因快速失活技术研究及应用[J].军事医学科学院院刊,2009,33(4):365-369. 被引量:1
  • 2张部昌,赵志虎,王以光,于秀琴,刘传暄,马清钧.糖多孢红霉菌A226的原生质体转化和染色体同源整合[J].生物技术通讯,2002,13(2):107-111. 被引量:20
  • 3[美]萨姆布鲁克(Sambrook,J·)等 著,金冬雁等.分子克隆实验指南[M]科学出版社,1992.
  • 4Hopwood DA.Soil to genomics:the Streptomyces chromosome. Annual Review of Genetics . 2006
  • 5Hengst C,Tran N,Bibb Me,t al.Genes essential for morphologicaldevelopment and antibiotic production in Streptomyces coelicolor aretargets of BldD during vegetative growth. Molecular Microbiology . 2010
  • 6Han S,Song P,Huang XDe,t al.7</sub>040,a member of TetR family related to the morphological differentia-tion of Saccharopolyspora erythraea&amp;sid=Current Microbiology&amp;aufirst=Han S');&#xA; ">Identification of SACE<sub>7</sub>040,a member of TetR family related to the morphological differentia-tion of Saccharopolyspora erythraea. Current Microbiology . 2011
  • 7Oliynyk M,Samborskyy M,Lester Je,t al.Complete genome se-quence of the erythromycin-producing bacterium Saccharopolysporaerythraea NRRL23338. Nature Biotechnology . 2007
  • 8Hopwood DA,Bibb MJ,Chater KF,et al.Genetic manipulation of stretomyces: a laboratory manual. . 1985
  • 9Summers R G,Donadio S,Staver M J,et al.Sequencing and mutagenesis of genes from the erythromycin biosynthetic gene cluster of Saccharopolyspora erythraea that are involved in L-mycarose and D-desosamine production. Microbiology . 1997
  • 10Yamamoto H,Maurer K H,Hutchinson C R.Transformation of streptomyces erythraeus. Journal of Antibiotics . 1986

二级参考文献34

  • 1Oliynyk M,Samborskyy M,Lester JB,et al.Complete genome sequence of the erythromycin-producing bacterium Saccharopolyspora erythraea NRRL23338[J].Nat Biotechnol,2007,25(4):447-453.
  • 2Menzella HG,Reeves CD.Combinatorial biosynthesis for drug development[J].Curr Opin Microbiol,2007,10(3):238-245.
  • 3MacNeil DJ,Gewain KM,Ruby CL,et al.Analysis of Streptomyces avermilis genes required for avermectin biosynthesis utilizing a novel integration vector[J].Gene,1992,111(1):61-68.
  • 4Hopwood DA,Bibb MJ,Chater KF,et al.Genetic manipulation of Streptomyces:a laboratory manual[M].Norwich:The John Innes Foundation,1985.
  • 5Yamamoto H,Maurer KH,Hutchinson CR.Transformation of streptomyces erythraeus[J].J Antibiot,1986,39(9):1304-1313.
  • 6Summers RG,Donadio S,Staver MJ,et al.Sequencing and mutagenesis of genes from the erythromycin biosynthetic gene cluster of Saccharopolyspora erythraea that are involved in L-mycarose and D-desosamine production[J].Microbiology,1997,143 (10):3251-3262.
  • 7Chng C,Lum AM,Vroom JA,et al.A key developmental regulator controls the synthesis of the antibiotic erythromycin in Saccharopolyspora erythraea[J].Proc Natl Acad Sci USA,2008,105(32):11346-11351.
  • 8Sharan SK,Thomason LC,Kuznetsov SG,et al.Recombineering:a homologous recombination-based method of genetic engineering[J].Nat Protoc,2009,4(2):206-223.
  • 9Baudin A,Ozier-KalogeropouLos O,Denouel A,et al.A simple and efficient method for direct gene deletion in Saccharomyces cerevisiae[J].Nucleic Acids Res,1993,21(14):3329-3330.
  • 10Murphy KC,Campellone KG,Poteete AR.PCR-mediated gene replacement in Escherichia coli[J].Gene,2000,246(1-2):321-330.

共引文献19

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部