Transfer of plasmid pTO1 from<i>Escherichia coli</i>to various representatives of the order Actinomycetales by intergeneric conjugation

Voeykova, Tatyana; Emelyanova, L. K.; Tabakov, Vyacheslav; Mkrtumyan, Nora

doi:10.1111/j.1574-6968.1998.tb12977.x

Cited by 51 publications

(6 citation statements)

References 9 publications

(4 reference statements)

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“…Since then, this method has been successfully applied to several Streptomyces species [2–4] via methylation-deficient E. coli strains, such as strain ET12567 [5], as a DNA donor to avoid methylated DNA restriction systems of actinomycetes [6]. Several cloning vectors that could be transferred from E. coli to Streptomyces species by conjugation method have been constructed [7, 8]. Among them, pSET152 is a nonreplicative plasmid in Streptomyces that carries the attP site and integrase gene of φ C31 phage and consequently can integrate into the chromosomal attB site of the bacteriophage φ C31.…”

Section: Introductionmentioning

confidence: 99%

Integrative Gene Cloning and Expression System for Streptomyces sp. US 24 and Streptomyces sp. TN 58 Bioactive Molecule Producing Strains

Sioud

Aigle²,

Karray-Rebai

et al. 2009

BioMed Research International

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Streptomyces sp. US 24 and Streptomyces sp. TN 58, two strains producing interesting bioactive molecules, were successfully transformed using E. coli ET12567 (pUZ8002), as a conjugal donor, carrying the integrative plasmid pSET152. For the Streptomyces sp. US 24 strain, two copies of this plasmid were tandemly integrated in the chromosome, whereas for Streptomyces sp. TN 58, the integration was in single copy at the attB site. Plasmid pSET152 was inherited every time for all analysed Streptomyces sp. US 24 and Streptomyces sp. TN 58 exconjugants under nonselective conditions. The growth, morphological differentiation, and active molecules production of all studied pSET152 integrated exconjugants were identical to those of wild type strains. Consequently, conjugal transfer using pSET152 integration system is a suitable means of genes transfer and expression for both studied strains. To validate the above gene transfer system, the glucose isomerase gene (xylA) from Streptomyces sp. SK was expressed in strain Streptomyces sp. TN 58. Obtained results indicated that heterologous glucose isomerase could be expressed and folded effectively. Glucose isomerase activity of the constructed TN 58 recombinant strain is of about eighteenfold higher than that of the Streptomyces sp. SK strain. Such results are certainly of importance due to the potential use of improved strains in biotechnological process for the production of high-fructose syrup from starch.

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Section: Introductionmentioning

confidence: 99%

Integrative Gene Cloning and Expression System for Streptomyces sp. US 24 and Streptomyces sp. TN 58 Bioactive Molecule Producing Strains

Sioud

Aigle²,

Karray-Rebai

et al. 2009

BioMed Research International

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“…The authors of the previous study used S. hygroscopicus SIPI-KF and E. coli S17-1. E. coli S17-1 contains a chromosomally integrated derivative of RP4, which stimulates the integration of DNA from the donor strain into the recipient genome ( Simon et al, 1983 ; Voeykova et al, 1998 ; Kieser et al, 2000 ). Thus, the status of the Fn Cas12a system in Streptomyces may be influenced by different donor strains.…”

Section: Discussionmentioning

confidence: 99%

Efficient Multiplex Genome Editing in Streptomyces via Engineered CRISPR-Cas12a Systems

Zhang

Zhu

et al. 2020

Front. Bioeng. Biotechnol.

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Streptomyces strains produce a great number of valuable natural products. With the development of genome sequencing, a vast number of biosynthetic gene clusters with high potential for use in the discovery of valuable clinical drugs have been revealed. Therefore, emerging needs for tools to manipulate these biosynthetic pathways are presented. Although the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas 9) system has exhibited great capabilities for gene editing in multiple Streptomyces strains, it has failed to work in some newly discovered strains and some important industrial strains. Additionally, the protospacer adjacent motif (PAM) recognition scope of this system sometimes limits its applications for generating precise site mutations and insertions. Here, we developed three efficient CRISPR-FnCas12a systems for multiplex genome editing in several Streptomyces strains. Each system exhibited advantages for different applications. The CRISPR-FnCas12a1 system was efficiently applied in the industrial strain Streptomyces hygroscopicus, in which SpCas9 does not work well. The CRISPR-FnCas12a2 system was used to delete large fragments ranging from 21.4 to 128 kb. Additionally, the CRISPR-FnCas12a3 system employing the engineered FnCas12a mutant EP16, which recognizes a broad spectrum of PAM sequences, was used to precisely perform site mutations and insertions. The CRISPR-FnCas12a3 system addressed the limitation of TTN PAM recognition in Streptomyces strains with high GC contents. In summary, all the CRISPR-FnCas12a systems developed in this study are powerful tools for precise and multiplex genome editing in Streptomyces strains.

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“…Typically, intergeneric conjugation is used as a means of gene transfer into streptomycetes to accomplish the first step (22,54,55). Intergeneric conjugation has already been successfully applied to species of various genera, e.g., Streptomyces, Actinomadura, Amycolatopsis, Arthrobacter, Micromonospora, Nocardia, Rhodococcus, Kitasatospora, and Saccharopolyspora (56)(57)(58)(59)(60). The advantage of this strategy is that it allows for the design and construction of recombinant plasmids in E. coli prior to transfer to the recipient of interest.…”

Section: Discussionmentioning

confidence: 99%

An Efficient Method To Generate Gene Deletion Mutants of the Rapamycin-Producing Bacterium Streptomyces iranensis HM 35

Netzker

Schroeckh

Gregory

et al. 2016

Appl Environ Microbiol

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Streptomyces iranensis HM 35 is an alternative rapamycin producer to Streptomyces rapamycinicus. Targeted genetic modification of rapamycin-producing actinomycetes is a powerful tool for the directed production of rapamycin derivatives, and it has also revealed some key features of the molecular biology of rapamycin formation in S. rapamycinicus. The approach depends upon efficient conjugational plasmid transfer from Escherichia coli to Streptomyces, and the failure of this step has frustrated its application to Streptomyces iranensis HM 35. Here, by systematically optimizing the process of conjugational plasmid transfer, including screening of various media, and by defining optimal temperatures and concentrations of antibiotics and Ca 2؉ ions in the conjugation media, we have achieved exconjugant formation for each of a series of gene deletions in S. iranensis HM 35. Among them were rapK, which generates the starter unit for rapamycin biosynthesis, and hutF, encoding a histidine catabolizing enzyme. The protocol that we have developed may allow efficient generation of targeted gene knockout mutants of Streptomyces species that are genetically difficult to manipulate. IMPORTANCE The developed protocol of conjugational plasmid transfer from Escherichia coli to Streptomyces iranensis may allow efficient generation of targeted gene knockout mutants of other genetically difficult to manipulate, but valuable, Streptomyces species. Since the discovery of streptomycin in 1943 (1), streptomycetes have been shown to produce thousands of compounds with possibly beneficial features, e.g., antibiotics, immunosuppressants, or anticancer drugs. Actinomycete-derived metabolites comprise over two-thirds of all known antibiotic compounds (2), and recent genome sequencing programs revealed that their biosynthesis potential has been underestimated. In their 8-to 12-Mb genomes, approximately 20 to 30 gene clusters encode the biosynthesis of secondary metabolites (3-5). One of the most important Streptomyces-derived compounds is the mechanistic target of rapamycin (mTOR) inhibitor rapamycin (sirolimus), a "billion dollar molecule" and, after cyclosporine, the most widely used immunosuppressant of microbial origin (6). Rapamycin was previously reported as an antifungal antibiotic produced by Streptomyces hygroscopicus ATCC 29253 (7), later renamed Streptomyces rapamycinicus (8). The rapamycin gene cluster in this strain has been sequenced (9), the biosynthetic pathway has been extensively characterized (10-14), and engineering of the cluster has yielded an impressive range of bioactive modified rapamycins (rapalogs) (15, 16), some in multigram amounts. So far, two other rapamycin-producing species are known: the taxonomically closely related Streptomyces iranensis HM 35 (5, 17) and Actinoplanes sp. strain N902-109 (18). To elucidate the molecular biology of rapamycin formation in these strains and to further exploit the physiological and pharmacological capability of rapamycin derivatives, genetic manipulation of these altern...

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Transfer of plasmid pTO1 fromEscherichia colito various representatives of the order Actinomycetales by intergeneric conjugation

Cited by 51 publications

References 9 publications

Integrative Gene Cloning and Expression System for Streptomyces sp. US 24 and Streptomyces sp. TN 58 Bioactive Molecule Producing Strains

Integrative Gene Cloning and Expression System for Streptomyces sp. US 24 and Streptomyces sp. TN 58 Bioactive Molecule Producing Strains

Efficient Multiplex Genome Editing in Streptomyces via Engineered CRISPR-Cas12a Systems

An Efficient Method To Generate Gene Deletion Mutants of the Rapamycin-Producing Bacterium Streptomyces iranensis HM 35

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