Development and optimization of genetic toolboxes for a fast-growing cyanobacterium Synechococcus elongatus UTEX 2973

Li, Shubin; Sun, Tao; Xu, Chunxiao; Chen, Lei; Zhang, Weiwen

doi:10.1016/j.ymben.2018.06.002

Cited by 91 publications

(108 citation statements)

References 69 publications

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“…In-depth studies of natural competence have largely focused on bacterial pathogens 3,40-42 , so little is known about the process in other prokaryotic phyla. In cyanobacteria, some of the conserved T4PM and competence proteins had been shown to be essential for competence 10,43 . Our work experimentally extends this knowledge to encompass all proteins that are required for the process including those that could not be identified solely by bioinformatics 11 .…”

Section: Discussionmentioning

confidence: 99%

The circadian clock and darkness control natural competence in cyanobacteria

Taton

Erikson

Yang

et al. 2019

Preprint

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Natural genetic competence-based transformation contributed to the evolution of prokaryotes, including the cyanobacterial phylum that established oxygenic photosynthesis. The cyanobacterium Synechococcus elongatus is noted both as a model system for analyzing a prokaryotic circadian clock and for its facile, but poorly understood, natural competence. Here a genome-wide screen aimed at determining the genetic basis of competence in cyanobacteria identified all genes required for natural transformation in S. elongatus, including conserved Type IV pilus, competence-associated, and newly described genes, and revealed that the circadian clock controls the process. The findings uncover a daily program that determines the state of competence in S. elongatus and adapts to seasonal changes of day-length. Pilus biogenesis occurs daily in the morning, but competence is maximal upon the coincidence of circadian dusk and the onset of darkness. As in heterotrophic bacteria, where natural competence is conditionally regulated by nutritional or other stress, cyanobacterial competence is conditional and is tied to the daily cycle set by the cell’s most critical nutritional source, the Sun.

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Section: Discussionmentioning

confidence: 99%

The circadian clock and darkness control natural competence in cyanobacteria

Taton

Erikson

Yang

et al. 2019

Preprint

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“…Pcpc560 (Huang et al, 2010;Zhou et al, 2014;Englund et al, 2016;Ruffing et al, 2016;Ferreira et al, 2018;Li et al, 2018;Liu and Pakrasi, 2018 Ptrc (Huang et al, 2010;Guerrero et al, 2012;Oliver et al, 2013;Markley et al, 2015;Ferreira et al, 2018;Li et al, 2018) TetR (aTc Inducible) L03 (Huang and Lindblad, 2013;Zess et al, 2016;Ferreira et al, 2018) efficient transformation using conjugation and electroporation (compared to homologous recombination); (2) avoidance of the need for time-consuming segregation to achieve incorporation of the transgenic sequence in all copies of the genome; and (3) increased orthogonality of the heterologous sequences, meaning no insertion in genomic loci that may not be neutral. A hybrid strategy that combines the advantages of using a shuttle vector and the stability of genomic insertion involves using native cyanobacterial plasmids, either as insertion sites (Ng et al, 2015) or as true shuttle vectors after isolation and modification (Jin et al, 2018).…”

Section: Constitutivementioning

confidence: 99%

The Synthetic Biology Toolkit for Photosynthetic Microorganisms

et al. 2019

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“…The results for UTEX 2973 were unexpected, and to our knowledge no studies to date have directly compared these promoters in this strain. Recent work has examined the expression of β-galactosidase using promoters such as Pcpc560 and Ptrc in UTEX 2973 (32). Li et al (32) highlighted that different growth environments (e.g.…”

Section: Heterologous and Synthetic Promotersmentioning

confidence: 99%

“…Recent work has examined the expression of β-galactosidase using promoters such as Pcpc560 and Ptrc in UTEX 2973 (32). Li et al (32) highlighted that different growth environments (e.g. light levels) can have significant effects on protein expression.…”

Section: Heterologous and Synthetic Promotersmentioning

confidence: 99%

“…Although exciting progress is being made in developing effective transformation systems, cyanobacteria still lag behind in the field of synthetic biology compared to bacterial (heterotrophic), yeast and mammalian systems. Relatively few broad-host range genetic parts have been characterised, but many libraries of parts for constructing regulatory modules and circuits are starting to become available, albeit using different standards, which makes them difficult to combine (23)(24)(25)(26)(27)(28)(29)(30)(31)(32)(33)(34). One key challenge is clear:…”

Section: Introductionmentioning

confidence: 99%

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CyanoGate: A Golden Gate modular cloning suite for engineering cyanobacteria based on the plant MoClo syntax

Vasudevan

et al. 2018

Preprint

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A Golden Gate-based toolkit for engineering cyanobacteria 2 ABSTRACT Recent advances in synthetic biology research have been underpinned by an exponential increase in available genomic information and a proliferation of advanced DNA assembly tools. The adoption of plasmid vector assembly standards and parts libraries has greatly enhanced the reproducibility of research and exchange of parts between different labs and biological systems. However, a standardised Modular Cloning (MoClo) system is not yet available for cyanobacteria, which lag behind other prokaryotes in synthetic biology despite their huge potential in biotechnological applications. By building on the assembly library and syntax of the Plant Golden Gate MoClo kit, we have developed a versatile system called CyanoGate that unites cyanobacteria with plant and algal systems. We have generated a suite of parts and acceptor vectors for making i) marked/unmarked knock-outs or integrations using an integrative acceptor vector, and ii) transient multigene expression and repression systems using known and novel replicative vectors. We have tested and compared the CyanoGate system in the established model cyanobacterium Synechocystis sp. PCC 6803 and the more recently described fast-growing strain Synechococcus elongatus UTEX 2973. The system is publicly available and can be readily expanded to accommodate other standardised MoClo parts.

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Development and optimization of genetic toolboxes for a fast-growing cyanobacterium Synechococcus elongatus UTEX 2973

Cited by 91 publications

References 69 publications

The circadian clock and darkness control natural competence in cyanobacteria

The circadian clock and darkness control natural competence in cyanobacteria

The Synthetic Biology Toolkit for Photosynthetic Microorganisms

CyanoGate: A Golden Gate modular cloning suite for engineering cyanobacteria based on the plant MoClo syntax

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