[Objective]The aim was to discuss the actinomycete biodiversity of iron-mine tailings by phylogenetic analysis of 12 typical isolates. [Method]The genomic DNAs were extracted by phenol-chloroform method; phylogeny ana...[Objective]The aim was to discuss the actinomycete biodiversity of iron-mine tailings by phylogenetic analysis of 12 typical isolates. [Method]The genomic DNAs were extracted by phenol-chloroform method; phylogeny analysis was carried out based on 16S rDNA PCR amplification and sequencing. [Result]The results showed that all the 12 strains belong to the genus Streptomyces sharing 98.7%-99.9% similarities with their nearest known neighbors. [Conclusion]Streptomyces is the dominant culturable actinomycete group of iron mine tailings,in which there are many potential novel species.展开更多
[ Objective ] The aim of this study is to identify 6 marine fungi species by analyzing ITS nucleotide sequence, which had been primarily identified as penicillium based on morphological characteristics. [ Method ] The...[ Objective ] The aim of this study is to identify 6 marine fungi species by analyzing ITS nucleotide sequence, which had been primarily identified as penicillium based on morphological characteristics. [ Method ] The ITS regions of these species were cloned by molecule biology method and phylogenetic analyzed using ClustalX1.83 software. [ Result] The ITS regions of these species were sequenced, and phylogenetic analysis between the yield sequences and the ITS sequences assessed in GenBank showed that the 6 strains all belonged to penicillium. [ Condusion] The present study suggests ITS sequence analysis could not be used as an only proof, but it is a very useful supplementary tool for the classification and identification of marine peniciUium combined with morphological characteristics.展开更多
[Objective] The aim of this study was to investigate the preparation method and amplification system of antagonistic streptomyces DNA templates based on AFLP assays, and also provide a basis for the application of AFL...[Objective] The aim of this study was to investigate the preparation method and amplification system of antagonistic streptomyces DNA templates based on AFLP assays, and also provide a basis for the application of AFLP technology in the analysis of streptomyces or even actinomyces. [Method] The DNAs were extracted by the modified CTAB method and amplified by the Pst Ⅰ/Mse Ⅰ AFLP kit and its reaction system. The amplified products were analyzed by the denatured polyacrylamide gel electrophoresis. [Result] The genomic DNAs of ten antagonistic strains of Streptomyces were extracted and tested. The result of 0.8% agarose gel electrophoresis showed that the major DNA bands were clear without degradation and RNA residue, with the fragment sizes ranging from 37.64 to 40.86 Kb. By ultraviolet spectrophotometry, the OD260/OD280 values varying from 1.625 to 1.833 were obtained. Furthermore, the agarose gel electrophoresis of DNA products digested by Pst Ⅰ/Mse Ⅰ presented the dispersed fluorescent long band, which indicated that the enzymatic hydrolysis was fully carried out. The amplified bands of DNA templates by the screened three pairs of primers were clear with rich polymorphism. [Conclusion] The preparation method and amplification system of DNA template established in this study can be used in the AFLP analysis of Streptomyces.展开更多
Neomycins are a group of aminoglycoside antibiotics with both clinical and agricultural applications.To elucidate the regulatory mechanism of neomycin biosynthesis,we completed draft genome sequencing of a neomycin pr...Neomycins are a group of aminoglycoside antibiotics with both clinical and agricultural applications.To elucidate the regulatory mechanism of neomycin biosynthesis,we completed draft genome sequencing of a neomycin producer Streptomyces fradiae CGMCC 4.7387 from marine sediments,and the neomycin biosynthesis gene cluster was identified.Inactivation of the afsA-g gene encoding a γ-butyrolactone(GBL) synthase in S.fradiae CGMCC 4.7387 resulted in a significant decrease of neomycin production.Quantitative RT-PCR analysis revealed that the transcriptional level of neoR and the aphA-neoGH operon were reduced in the afsA-g::aac(3)Ⅳ mutant.Interestingly,a conserved binding site of AdpA,a key activator in the GBL regulatory cascade,was discovered upstream of neoR,a putative regulatory gene encoding a protein with an ATPase domain and a tetratricopeptide repeat domain.When neoR was inactivated,the neomycin production was reduced about 40%in comparison with the WT strain.Quantitative RT-PCR analysis revealed that the transcriptional levels of genes in the aphA-neoGH operon were reduced clearly in the neoR::aac(3)Ⅳ mutant.Finally,the titers of neomycin were improved considerably by overexpression of qfsA-gand neoR in S.fradiae CGMCC 4.7387.展开更多
基金Supported by National Natural Science Foundation of China(30800003)Provincial Foundation Advanced Project of Hebei Uni-versity (2006Y09)~~
文摘[Objective]The aim was to discuss the actinomycete biodiversity of iron-mine tailings by phylogenetic analysis of 12 typical isolates. [Method]The genomic DNAs were extracted by phenol-chloroform method; phylogeny analysis was carried out based on 16S rDNA PCR amplification and sequencing. [Result]The results showed that all the 12 strains belong to the genus Streptomyces sharing 98.7%-99.9% similarities with their nearest known neighbors. [Conclusion]Streptomyces is the dominant culturable actinomycete group of iron mine tailings,in which there are many potential novel species.
基金Supported by Standardization Arrangement and Integration and Shareof the Offshore Ocean Microorganism Resources(2004DKA30640)~~
文摘[ Objective ] The aim of this study is to identify 6 marine fungi species by analyzing ITS nucleotide sequence, which had been primarily identified as penicillium based on morphological characteristics. [ Method ] The ITS regions of these species were cloned by molecule biology method and phylogenetic analyzed using ClustalX1.83 software. [ Result] The ITS regions of these species were sequenced, and phylogenetic analysis between the yield sequences and the ITS sequences assessed in GenBank showed that the 6 strains all belonged to penicillium. [ Condusion] The present study suggests ITS sequence analysis could not be used as an only proof, but it is a very useful supplementary tool for the classification and identification of marine peniciUium combined with morphological characteristics.
基金Supported by the National Natural Science Foundation of China(30671346)Beijing Natural Science Foundation (6072010)Beijing Municipal Science and Technology Plan Project (D0706005000091)~~
文摘[Objective] The aim of this study was to investigate the preparation method and amplification system of antagonistic streptomyces DNA templates based on AFLP assays, and also provide a basis for the application of AFLP technology in the analysis of streptomyces or even actinomyces. [Method] The DNAs were extracted by the modified CTAB method and amplified by the Pst Ⅰ/Mse Ⅰ AFLP kit and its reaction system. The amplified products were analyzed by the denatured polyacrylamide gel electrophoresis. [Result] The genomic DNAs of ten antagonistic strains of Streptomyces were extracted and tested. The result of 0.8% agarose gel electrophoresis showed that the major DNA bands were clear without degradation and RNA residue, with the fragment sizes ranging from 37.64 to 40.86 Kb. By ultraviolet spectrophotometry, the OD260/OD280 values varying from 1.625 to 1.833 were obtained. Furthermore, the agarose gel electrophoresis of DNA products digested by Pst Ⅰ/Mse Ⅰ presented the dispersed fluorescent long band, which indicated that the enzymatic hydrolysis was fully carried out. The amplified bands of DNA templates by the screened three pairs of primers were clear with rich polymorphism. [Conclusion] The preparation method and amplification system of DNA template established in this study can be used in the AFLP analysis of Streptomyces.
基金funded in part by the Ministry of Science and Technology of China(2015CB150600)the National Natural Science Foundation of China(31370095 and 31522001)
文摘Neomycins are a group of aminoglycoside antibiotics with both clinical and agricultural applications.To elucidate the regulatory mechanism of neomycin biosynthesis,we completed draft genome sequencing of a neomycin producer Streptomyces fradiae CGMCC 4.7387 from marine sediments,and the neomycin biosynthesis gene cluster was identified.Inactivation of the afsA-g gene encoding a γ-butyrolactone(GBL) synthase in S.fradiae CGMCC 4.7387 resulted in a significant decrease of neomycin production.Quantitative RT-PCR analysis revealed that the transcriptional level of neoR and the aphA-neoGH operon were reduced in the afsA-g::aac(3)Ⅳ mutant.Interestingly,a conserved binding site of AdpA,a key activator in the GBL regulatory cascade,was discovered upstream of neoR,a putative regulatory gene encoding a protein with an ATPase domain and a tetratricopeptide repeat domain.When neoR was inactivated,the neomycin production was reduced about 40%in comparison with the WT strain.Quantitative RT-PCR analysis revealed that the transcriptional levels of genes in the aphA-neoGH operon were reduced clearly in the neoR::aac(3)Ⅳ mutant.Finally,the titers of neomycin were improved considerably by overexpression of qfsA-gand neoR in S.fradiae CGMCC 4.7387.
