Laboratory Maintenance of <i>Streptomyces</i> Species

Shepherd, Micah D.; Kharel, Madan K.; Bosserman, Mary A.; Rohr, Jürgen

doi:10.1002/9780471729259.mc10e01s18

Cited by 74 publications

(51 citation statements)

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“…E. coli bacterial cultures were grown in Luria-Bertani (LB) Miller medium (BD Biosciences, USA) at 37 °C. S. venezuelae ATCC 10712 was grown on MYM media (28), and sporulated on mannitol soy (MS) agar media supplemented with 10 mM magnesium chloride (28). Cultures were supplemented with carbenicillin (100 mg/L, Sigma Aldrich, USA), kanamycin (50 mg/L, Sigma Aldrich, USA), or spectinomycin (100 mg/L for E. coli or 400 mg/L for S. venezuelae , Sigma Aldrich, USA) All bacterial strains and plasmids used in this work are listed in Table 2 and are available through the public instance of the JBEI registry.…”

Section: Methodsmentioning

confidence: 99%

Characterization of an ECF56-family sigma factor fromStreptomyces venezuelaereveals a highly conserved regulome

Thompson

Zargar

Cruz-Morales

et al. 2020

Preprint

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34Bacteria often possess alternative sigma factors that initiate the transcription of 35 specific genes under environmental stresses, the largest and most diverse group being 36 the extracytoplasmic function (ECF) sigma factors. The regulation of ECF activity is 37 crucial for ensuring the distinct transcription of stress responsive genes only occurs 38 under the appropriate conditions. While most ECFs are comprised of only the core σ 2 39 and σ 4 regions, a unique form of ECF sigma factor regulation also contains a C-terminal 40 extension bearing homology to the NTF2 superfamily of protein domains. While 41 previous work has shown that this NTF2 domain can affect transcriptional activity in vivo 42 in ECF41 and ECF42, its role in the newly classified ECF56 subgroup is unknown. In 43 this work, we show that truncation of the C-terminus of the ECF56 sigma factor 44 SVEN_4562 of Streptomyces venezuelae upregulates its activity in a hybrid assay. 45 Through transcriptomics in S. venezuelae, we found that this truncated ECF56 sigma 46 factor has a highly conserved promoter sequence in vivo. Bioinformatic assays 47 ECF56 family of sigma factors is highly conserved and performs important functions yet 53 to be understood in Actinobacteria. 54 55 Importance 56 Most ECF sigma-factors rely on anti-sigma factor regulation; in contrast, the 57 unique classes of ECF sigma-factors that contain a C-terminal extension are thought to 58 respond directly to an environmental signal. Here we show that the cis-acting regulatory 59 element of the ECF56 regulon is likely highly conserved in many Actinobacteria, with 60 exact nucleotide level conservation over ~2 billion years of evolution. The high 61 conservation of this genetic architecture, as well as a conserved gene content within the 62 regulon, strongly point to a specialized and important role in Actinobacteria biology. 63 64

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Section: Methodsmentioning

confidence: 99%

Characterization of an ECF56-family sigma factor fromStreptomyces venezuelaereveals a highly conserved regulome

Thompson

Zargar

Cruz-Morales

et al. 2020

Preprint

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“…[225] Dichlormethan [316] die Sporen wurden anschließend geerntet und in Glycerol gelagert. [317] Hemmhoftest Eine S. coelicolor-Sporenlösung (100 µL, 1×10 8 aktive Sporen/ml) wurde auf Petrischalen gleichmäßig verteilt (8.5 cm) die 30 mL Minimal-Medium enthielten. [318] Anschließend wurden mit der Rückseite einer Pasteur-Pipette Löcher in das Medium gestanzt und mit 100 µL der Analytlösung in DMSO gefüllt.…”

Section: Synthese Der Nicht Fluorierten Biscarboxamid-ligandenunclassified

Totalsynthese von Nosiheptid

Wojtas

Riedrich

et al. 2016

Angewandte Chemie

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Eine Totalsynthese des bismakrocyclischen Thiopeptidantibiotikums Nosiheptid gelang durch den Aufbau einer voll funktionalisierten linearen Vorstufe und konsekutive Makrocyclisierungen. Schlüsselmerkmale waren eine kritische Makrothiolactonisierung und eine milde Entschützungsstrategie für den 3‐Hydroxypyridin‐Kern. Der Naturstoff war mit isoliertem authentischem Material im Hinblick auf spektroskopische Daten und antimikrobielle Aktivität identisch.

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“…These isolates were routinely maintained onto starch nitrate agar and some selected isolates were subject for long term preservation in slant medium or as spores glycerol stocks (Kieser et al, 2000;Shepherd et al, 2010).…”

Section: Streptomyces Isolatesmentioning

confidence: 99%