Construction of a promoter‐probe vector for the methanol‐utilizing bacterium acetobacter methanolicus MB 58

Schröder, R; Engel, J.; Chistoserdov, A. Y.; Tsygankov, Y. D.

doi:10.1002/abio.370090306

Cited by 3 publications

(4 citation statements)

References 16 publications

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“…As described before, the AAB genus Acidomonas was established since several characteristics of the former species Acetobacter methanolicus could be distinguished from other type and representative AAB including Acetobacter strains, resulting in the renaming of A. methanolicus to Acidomonas methanolica (Urakami et al 1989 ). For Acidomonas expression vectors including a basic promoter probe vector have been created based on the popular broad-host-range multi-copy plasmid RSF1010 (Schröder et al 1989 ; Schröder et al 1991 ). In A. methanolica , a RSF1010-based derivative was used to express the core antigen gene of hepatitis B virus subtype ayw under control of the E. coli P lac promoter alone and under tandem control of the strong promoter P acm from the Acetobacter phage Acm l and from P lac .…”

Section: Target Gene Expression In Acidomonasmentioning

confidence: 99%

On the way toward regulatable expression systems in acetic acid bacteria: target gene expression and use cases

Fricke

Klemm

Bott

et al. 2021

Appl Microbiol Biotechnol

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Acetic acid bacteria (AAB) are valuable biocatalysts for which there is growing interest in understanding their basics including physiology and biochemistry. This is accompanied by growing demands for metabolic engineering of AAB to take advantage of their properties and to improve their biomanufacturing efficiencies. Controlled expression of target genes is key to fundamental and applied microbiological research. In order to get an overview of expression systems and their applications in AAB, we carried out a comprehensive literature search using the Web of Science Core Collection database. The Acetobacteraceae family currently comprises 49 genera. We found overall 6097 publications related to one or more AAB genera since 1973, when the first successful recombinant DNA experiments in Escherichia coli have been published. The use of plasmids in AAB began in 1985 and till today was reported for only nine out of the 49 AAB genera currently described. We found at least five major expression plasmid lineages and a multitude of further expression plasmids, almost all enabling only constitutive target gene expression. Only recently, two regulatable expression systems became available for AAB, an N-acyl homoserine lactone (AHL)-inducible system for Komagataeibacter rhaeticus and an l-arabinose-inducible system for Gluconobacter oxydans. Thus, after 35 years of constitutive target gene expression in AAB, we now have the first regulatable expression systems for AAB in hand and further regulatable expression systems for AAB can be expected. Key points • Literature search revealed developments and usage of expression systems in AAB. • Only recently 2 regulatable plasmid systems became available for only 2 AAB genera. • Further regulatable expression systems for AAB are in sight.

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Section: Target Gene Expression In Acidomonasmentioning

confidence: 99%

On the way toward regulatable expression systems in acetic acid bacteria: target gene expression and use cases

Fricke

Klemm

Bott

et al. 2021

Appl Microbiol Biotechnol

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“…3. In comparison with the plasmid pRCF1, which contains only the promoter of the cloned DNA fragment, the specific homoserine dehydrogenase activity increased by 2 to 3 fold in the plasmid pRCFtac containing the tac promoter in the tandem position and by 5 fold in the plasmid pRCF119 containing the acml phage promoter.…”

Section: Transfer and Expression Of The Ptlb Gene In E Coli And A mentioning

confidence: 99%

“…These plasmids can be transferred to and be stably maintained in many gram-negative bacteria, including the acidophilic methanol-assimilating bacterium A. methanolicus. One of the most popular plasmids used for this purpose is the multicopy number plasmid RSFlOlO [2], which can be mobilized into A. methanolicus by a triparental mating technique using pRK2013 as a helper plasmid [3]. An optimal expression of a foreign gene in a given host requires an optimal initiation of transcription and translation.…”

Section: Introductionmentioning

confidence: 99%

Construction of broad‐host‐range plasmids for the expression of heterologous genes in Acetobacter methanolicus B58

Föllner

Schröder

Babel

1994

Acta Biotechnologica

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The construction of different plasmids reported here on the basis of a broad-host-range RSFlOlO replicon allows an efficient expression of heterologous genes in the acidophilic methanol-assimilating bacterium Acetobacter methanolicus B58. The promoter-probe vector pRS2Ol was used for the identification and isolation of the promoter containing sequences derived from the DNA of the Acerobacrer phage Acml. Further, this plasmid was coupled with the Escherichia coli promoters tac and pr creating the expression vectors pRS20ltac and pRSSOlp,, respectively. After the insertion of the chloramphenicol acetyltransferase (cat) gene of the cloned promoters downstream, the chloramphenicol acetyltransferase (CAT) was determined in a cell-free extract of both E. coli and A. methanolicus. Using E. coli promoters as well as promoters of the Acetobacter phage Acml arranged in tandem with the promoters of the heterologous genes to be expressed, the pectat lyase gene @rm) of Erwinia carotouora and the threonine A gene (thrA) of E. coli were successfully expressed in A. rnethanolicus. The stability of recombinant plasmids under various conditions in A. methanolicus strains was tested using antibiotic-free media.

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“…In previous work it was shown that the broad-host-range plasmid RSF1010 and its derivatives can be mobilized and stably maintained in this methylotrophic bacterium (SchrSder et al 1989). Therefore, constructed plasmids derived from RSF1010 were transferred via mobilization into the new host using pRK2013 as helper plasmid.…”

Section: Expression O F the Core Antigen Gene O F H B V In E Coli Anmentioning

confidence: 99%

Expression of the core antigen gene of hepatitis B virus (HBV) in Acetobacter methanolicus using broad-host-range vectors

Schröder

Maaßen

Lippoldt

et al. 1991

Appl Microbiol Biotechnol

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Using the broad-host-range promoter probe vector pRS201 for cloning of phage Acm1 promoters, we established a convenient vector system for expression of heterologous genes in different Gram-negative bacteria. The usefulness of this system was demonstrated by expression of the HBV core gene in Acetobacter methanolicus. Plasmids carrying the HBV core gene downstream of different Acm1-phage promoters were transferred to A. methanolicus, a new potential host for recombinant DNA expression. Using enzyme immunoassay and immunoblot techniques, the amount and composition of core antigen produced in A. methanolicus were compared with that derived from Escherichia coli. The expression of immunoreactive core antigen in A. methanolicus exceeds by sevenfold that in E. coli using an expression system with tandemly arranged promoters. Morphological observations by electron microscopy show that the HBV core gene products isolated from both hosts are assembled into regular spherical particles with a diameter of about 28 nm that are comparable to original viral nucleocapsids.

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Construction of a promoter‐probe vector for the methanol‐utilizing bacterium acetobacter methanolicus MB 58

Cited by 3 publications

References 16 publications

On the way toward regulatable expression systems in acetic acid bacteria: target gene expression and use cases

On the way toward regulatable expression systems in acetic acid bacteria: target gene expression and use cases

Construction of broad‐host‐range plasmids for the expression of heterologous genes in Acetobacter methanolicus B58

Expression of the core antigen gene of hepatitis B virus (HBV) in Acetobacter methanolicus using broad-host-range vectors

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