Evidence map›Paper›PMID 32070286›Full record

ArticleBMC microbiology2020

Identification of promoter elements in the Dolichospermum circinale AWQC131C saxitoxin gene cluster and the experimental analysis of their use for heterologous expression.

Paul M D'Agostino, Bakir Al-Sinawi, Rabia Mazmouz, Julia Muenchhoff, Brett A Neilan, Michelle C Moffitt

Open access · goldAbstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in BMC microbiology, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.

0numbers the graph read from it
0cells of the map it votes in
5citing papers in PubMed
0.3field-weighted citation impact, top 40% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

5 citing papers in PubMed, 8 citations in OpenAlex.

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4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

6 authors at 2 institutions in 2 countries.

Paul M D'AgostinoSchool of Science, Western Sydney University, Sydney, NSW, Australia.ORCID 0000-0002-8323-5416
Bakir Al-SinawiSchool of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW, Australia.
Rabia MazmouzSchool of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW, Australia.
Julia MuenchhoffSchool of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW, Australia.
Brett A NeilanSchool of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW, Australia. brett.neilan@newcastle.edu.au.
Michelle C MoffittSchool of Science, Western Sydney University, Sydney, NSW, Australia. m.moffitt@westernsydney.edu.au.ORCID 0000-0001-9872-3175
UNSW Sydney · AUWestern Sydney University · AU

Funding

Australian Research Council Discovery Project DP0880264
6 · The paper itself

Abstract

backgroundDolichospermum circinale is a filamentous bloom-forming cyanobacterium responsible for biosynthesis of the paralytic shellfish toxins (PST), including saxitoxin. PSTs are neurotoxins and in their purified form are important analytical standards for monitoring the quality of water and seafood and biomedical research tools for studying neuronal sodium channels. More recently, PSTs have been recognised for their utility as local anaesthetics. Characterisation of the transcriptional elements within the saxitoxin (sxt) biosynthetic gene cluster (BGC) is a first step towards accessing these molecules for biotechnology.

resultsIn D. circinale AWQC131C the sxt BGC is transcribed from two bidirectional promoter regions encoding five individual promoters. These promoters were identified experimentally using 5' RACE and their activity assessed via coupling to a lux reporter system in E. coli and Synechocystis sp. PCC 6803. Transcription of the predicted drug/metabolite transporter (DMT) encoded by sxtPER was found to initiate from two promoters, PsxtPER1 and PsxtPER2. In E. coli, strong expression of lux from PsxtP, PsxtD and PsxtPER1 was observed while expression from Porf24 and PsxtPER2 was remarkably weaker. In contrast, heterologous expression in Synechocystis sp. PCC 6803 showed that expression of lux from PsxtP, PsxtPER1, and Porf24 promoters was statistically higher compared to the non-promoter control, while PsxtD showed poor activity under the described conditions.

conclusionsBoth of the heterologous hosts investigated in this study exhibited high expression levels from three of the five sxt promoters. These results indicate that the majority of the native sxt promoters appear active in different heterologous hosts, simplifying initial cloning efforts. Therefore, heterologous expression of the sxt BGC in either E. coli or Synechocystis could be a viable first option for producing PSTs for industrial or biomedical purposes.

Indexed as

Bacterial ProteinsCyanobacteriaModels, GeneticMultigene FamilyPromoter Regions, GeneticSaxitoxinBacterial ProteinsSaxitoxinAnabaena circinalisCyanobacteriaDolichospermum circinaleHeterologous expressionParalytic shellfish toxinsPromoterRegulationSaxitoxinTranscription

Identifiers

PMID32070286
PMCPMC7027233
OpenAlexW3012896614

What OpenQuestion holds

Textmetadata
LicenceCC BY
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.