PCR primers targeting the 16S rRNA gene for the specific detection of streptomycetes

Rintala, Helena; Nevalainen, Aino; Rönkä, Elina; Suutari, Merja

doi:10.1006/mcpr.2001.0379

Cited by 70 publications

(65 citation statements)

References 27 publications

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“…The primers were positively used to amplify genomic DNA from the isolated samples. These results are in agreement with Rintala et al [30], who showed that these primers are specific for Streptomyces.…”

Section: Pcr Amplification and Primer Specificitysupporting

confidence: 93%

“…The sequencing Streptomyces-specific PCR primers used were StrepB, 5'-ACA AGC CCT GGA AAC GGG T-3' (forward) and StrepF 5'-ACG TGT GCA GCC CAA GACA-3' (reveres) using Biolego BV software (Biolegio, Nijmegen, the Netherlands). These primers were used for the amplification of 16S rRNA fragments from genomic DNA isolated Streptomyces [30].…”

Section: Polymerase Chain Reaction (Pcr) Amplification and Sequencingmentioning

confidence: 99%

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MOLECULAR AND MORPHOLOGICAL IDENTIFICATION OF STREPTOMYCES SP. NRC-88 NOVA SPECIES AS Β-Lactamase INHIBITOR FOR PHARMACEUTICAL APPLICATION

Awad

Germoush

2017

Asian J Pharm Clin Res

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Objective: Clavulanic acid (CA) is a vital agent in the treatment of bacterial infections since it is a potent inhibitor of an extensive variety of β-lactamase enzymes. Its production from Streptomyces strains is fact of expanding clinical significance. This study aimed to isolate and characterize a promising Streptomyces (S) species strain which produced an effective β-lactamase inhibitor.Methods: Streptomyces sp. NRC-88 was isolated from an Egyptian soil sample. The phenotypic and phylogenetic examinations of 16S rRNA gene were investigated. The active metabolite of this strain (CA) was determined by particular synergistic bioassay, spectrophotometric assay, recognized by thin layer chromatography, and structure of the CA affirmed by high-performance liquid chromatography (HPLC) method. Results:A phylogenetic examination of the 16S rRNA gene sequence of the NRC-88 strain consistent with conventional taxonomy was carried out, and confirmed that the strain NRC-88 was most similar to S. aburaviensis S-66 (99%). The active metabolite of this strain (CA) was determined by different methods and confirmed the structure of the CA by the HPLC method. It produced up to 87 mg/l in a specific CA production medium. Conclusion:A new species of Streptomyces sp. NRC-88 isolated and recognized by phenotypic and genotypic proof. This strain suggested the name, Streptomyces sp. NRC-88 (accession number KM014489). It was able to produce CA as the β-lactamase inhibitor.

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Section: Pcr Amplification and Primer Specificitysupporting

confidence: 93%

Section: Polymerase Chain Reaction (Pcr) Amplification and Sequencingmentioning

confidence: 99%

MOLECULAR AND MORPHOLOGICAL IDENTIFICATION OF STREPTOMYCES SP. NRC-88 NOVA SPECIES AS Β-Lactamase INHIBITOR FOR PHARMACEUTICAL APPLICATION

Awad

Germoush

2017

Asian J Pharm Clin Res

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“…DNA was extracted from whole beewolf antennae according to a standard phenol/chloroform extraction protocol (Sambrook et al, 1989). The following primer pairs were used for the amplification of Streptomyces 16S rRNA gene: fD1 (forward) (Weisburg et al, 1991) and StrepF (reverse) (Rintala et al, 2001), Act-S20 (forward) (Stach et al, 2003) and rP2 (reverse) (Weisburg et al, 1991). While primers fD1 and rP2 can be used to amplify a wide range of eubacterial 16S rRNA, the combination with StrepF and Act-S20 ensured that the PCR was specific for actinomycete 16S rRNA.…”

Section: Methodsmentioning

confidence: 99%

‘Candidatus Streptomyces philanthi’, an endosymbiotic streptomycete in the antennae of Philanthus digger wasps

Kaltenpoth

Goettler

Dale

et al. 2006

International Journal of Systematic and Evolutionary Microbiology

126

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Symbiotic interactions with bacteria are essential for the survival and reproduction of many insects. The European beewolf (Philanthus triangulum, Hymenoptera, Crabronidae) engages in a highly specific association with bacteria of the genus Streptomyces that appears to protect beewolf offspring against infection by pathogens. Using transmission and scanning electron microscopy, the bacteria were located in the antennal glands of female wasps, where they form dense cell clusters. Using genetic methods, closely related streptomycetes were found in the antennae of 27 Philanthus species (including two subspecies of P. triangulum from distant localities). In contrast, no endosymbionts could be detected in the antennae of other genera within the subfamily Philanthinae (Aphilanthops, Clypeadon and Cerceris). On the basis of morphological, genetic and ecological data, ‘Candidatus Streptomyces philanthi’ is proposed. 16S rRNA gene sequence data are provided for 28 ecotypes of ‘Candidatus Streptomyces philanthi’ that reside in different host species and subspecies of the genus Philanthus. Primers for the selective amplification of ‘Candidatus Streptomyces philanthi’ and an oligonucleotide probe for specific detection by fluorescence in situ hybridization (FISH) are described.

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“…The "Rev" primers acted as primers for the reverse transcription. The oligonucleotides used for amplification of the 16 S rRNA gene from Streptomyces species were previously reported (29).…”

Section: Rna Isolation and Analysis Of Transcriptsmentioning

confidence: 99%

Iron-mediated Oxidation Induces Conformational Changes within the Redox-sensing Protein HbpS

Lucana

Roscher

Honigmann

et al. 2010

Journal of Biological Chemistry

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HbpS is an extracellular oligomeric protein, which has been shown to act in concert with the two-component system SenSSenR during the sensing of redox stress. HbpS can bind and degrade heme under oxidative stress conditions, leading to a free iron ion. The liberated iron is subsequently coordinated on the protein surface. Furthermore, HbpS has been shown to modulate the phosphorylation state of the sensor kinase SenS as, in the absence of oxidative stress conditions, HbpS inhibits SenS autophosphorylation whereas the presence of heme or iron ions and redox-stressing agents enhances it. Using HbpS wild type and mutants as well as different biochemical and biophysical approaches, we show that iron-mediated oxidative stress induces both secondary structure and overall intrinsic conformational changes within HbpS. We demonstrate in addition that HbpS is oxidatively modified, leading to the generation of highly reactive carbonyl groups and tyrosine-tyrosine bonds. Further examination of the crystal structure and subsequent mutational analyses allowed the identification of the tyrosine residue participating in dityrosine formation, which occurs between two monomers within the octomeric assembly. Therefore, it is proposed that oxidative modifications causing structural and conformational changes are responsible for the control of SenS and hence of the HbpS-SenS-SenR signaling cascade.Iron is the fourth most abundant element in the Earth's crust and is an essential trace mineral for nearly all known organisms. Under physiological conditions, it exists either in the reduced Fe 2ϩ (ferrous) or in the oxidized Fe 3ϩ (ferric) form. It plays a crucial role in many biological processes, as photosynthesis, N 2 fixation, H 2 production and consumption, respiration, oxygen transport, or gene regulation. Because of its redox potential ranging from Ϫ300 to ϩ700 mV, iron is a versatile prosthetic component that can be incorporated into proteins either as a mono-or binuclear species, or in a more complex form as part of iron-sulfur clusters or heme groups (1-3).In the presence of oxygen, iron ions frequently lead to the formation of redox stress by the generation of reactive oxygen species ( ROS can provoke the damage of DNA, lipids, and proteins (4 -6). For instance, when proteins are exposed to ROS, they undergo a variety of oxidative modifications including: Nitration of aromatic amino acid residues, hydroxylation of aromatic groups, and aliphatic amino acid side-chains, sulfoxidation of methionine residues, and conversion of some amino acid residues to carbonyl derivatives. Oxidation can also induce the cleavage of the polypeptide chain and the formation of crosslinked protein derivatives (7,8). These modifications can lead to functional changes of proteins that subsequently disturb the cellular metabolism. Thus, while bacteria and other organisms have to ensure that enough iron ions are present for the diverse biochemical reactions, they also have to avoid their harmful effects.We have previously identified the two-component sy...

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PCR primers targeting the 16S rRNA gene for the specific detection of streptomycetes

Cited by 70 publications

References 27 publications

MOLECULAR AND MORPHOLOGICAL IDENTIFICATION OF STREPTOMYCES SP. NRC-88 NOVA SPECIES AS Β-Lactamase INHIBITOR FOR PHARMACEUTICAL APPLICATION

MOLECULAR AND MORPHOLOGICAL IDENTIFICATION OF STREPTOMYCES SP. NRC-88 NOVA SPECIES AS Β-Lactamase INHIBITOR FOR PHARMACEUTICAL APPLICATION

‘Candidatus Streptomyces philanthi’, an endosymbiotic streptomycete in the antennae of Philanthus digger wasps

Iron-mediated Oxidation Induces Conformational Changes within the Redox-sensing Protein HbpS

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