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Claudia Argueta

Bio: Claudia Argueta is an academic researcher from California State University, Northridge. The author has contributed to research in topics: Hormogonium & Nostoc punctiforme. The author has an hindex of 3, co-authored 3 publications receiving 90 citations.

Papers
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Journal ArticleDOI
TL;DR: Two transcriptional reporter shuttle vectors constructed for the filamentous cyanobacterium Nostoc punctiforme using the green fluorescence protein (GFP) reporter demonstrate the utility of these GFP vectors to study cell-type-specific gene expression in differentiating filamentous Cyanobacteria.

43 citations

Journal ArticleDOI
TL;DR: To identify genes associated with akinete development, differential display was used to amplify and compare cDNA from a wild‐type and zwf mutant strain of N. punctiforme following a switch to dark heterotrophic conditions and three novel akinete‐expressed genes were identified.
Abstract: Akinetes are spore-like resting cells formed by certain filamentous cyanobacteria that have increased resistance to environmental stress. They can be found at low frequencies in dense cultures experiencing low light or phosphate limitation, but also form at high frequencies in a zwf mutant strain of Nostoc punctiforme following dark incubation in the presence of fructose. The wild-type strain is capable of facultative heterotrophic growth under these conditions and does not form akinetes. To identify genes associated with akinete development, differential display was used to amplify and compare cDNA from a wild-type and zwf mutant strain of N. punctiforme following a switch to dark heterotrophic conditions. Screening of candidate genes by reverse transcriptase real-time quantitative PCR and subsequent testing for akinete-specific expression using GFP transcriptional reporter plasmids lead to the identification of three novel akinete-expressed genes. The genes identified from the screening encoded for proteins homologous to an aminopeptidase (aapN), a zinc protease (hap) and an ATP-binding cassette (ABC)-type transporter (aet). Expression of hap was also increased in developing hormogonia, a transient type of differentiated filament capable of gliding motility. Transcriptional start sites for akinete-expressed genes were determined using random amplification of cDNA ends (RACE), and promoter regions were compared with orthologues in other filamentous cyanobacteria to identify putative regulatory sequences.

30 citations

Journal ArticleDOI
TL;DR: P phenotypic and genetic evidence showing near-synchronous induction of dark-induced zwf akinetes indicates that this system will provide a valuable tool for the molecular genetic study of akinete development in N. punctiforme.
Abstract: Nostoc punctiforme is a filamentous cyanobacterium that is capable of dark heterotrophy and cellular differentiation into nitrogen-fixing heterocysts, motile hormogonia, or spore-like akinetes. The study of akinete differentiation at the molecular level has been limited by the asynchronous development and limited number of akinetes formed within a filament. A system in which to study the development and genetic regulation of akinetes was investigated using a zwf mutant lacking glucose-6-phosphate dehydrogenase, the initial enzyme of the oxidative pentose phosphate pathway. Upon dark incubation in the presence of fructose, the zwf− strain ceased growth and differentiated into akinete-like cells, whereas the wild-type strain exhibited heterotrophic growth. Dark-induced zwf akinetes exhibited periodic acid–Schiff staining characteristics identical to that observed for wild-type akinetes, and synchronous induction of akinetes occurred in treated cultures. Dark-induced zwf akinetes exhibited increased resistance to the environmental stresses of desiccation, cold, or treatment with lysozyme relative to vegetative cells of both strains. Transcription of the avaK akinete marker gene was strongly induced in developing zwf akinetes as shown by Northern blotting and green fluorescent protein transcriptional reporter fusions. ATP levels did not vary significantly between dark incubated strains, indicating that a signal other than energy level may trigger akinete formation. This phenotypic and genetic evidence showing near-synchronous induction of dark-induced zwf akinetes indicates that this system will provide a valuable tool for the molecular genetic study of akinete development in N. punctiforme.

26 citations


Cited by
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Journal ArticleDOI
TL;DR: This Review addresses cyanob bacterial intercellular communication, the supracellular structure of the cyanobacterial filament and the basic principles that govern the process of heterocyst differentiation.
Abstract: Within the wide biodiversity that is found in the bacterial world, Cyanobacteria represents a unique phylogenetic group that is responsible for a key metabolic process in the biosphere - oxygenic photosynthesis - and that includes representatives exhibiting complex morphologies. Many cyanobacteria are multicellular, growing as filaments of cells in which some cells can differentiate to carry out specialized functions. These differentiated cells include resistance and dispersal forms as well as a metabolically specialized form that is devoted to N(2) fixation, known as the heterocyst. In this Review we address cyanobacterial intercellular communication, the supracellular structure of the cyanobacterial filament and the basic principles that govern the process of heterocyst differentiation.

400 citations

Book ChapterDOI
TL;DR: With this chapter, the synthetic biologist is given an overview of existing biological parts, tools and protocols for the genetic engineering, and molecular analysis of cyanobacteria for Synthetic Biology applications.
Abstract: Cyanobacteria are solar-powered cell factories that can be engineered to supply us with renewable fuels and chemicals. To do so robust and well-working biological parts and tools are necessary. Parts for controlling gene expression are of special importance in living systems, and specifically promoters are needed for enabling and simplifying rational design. Synthetic biology is an engineering science that incorporates principles such as decoupling, standardization and modularity to enable the design and construction of more advanced systems from simpler parts and the re-use of parts in new contexts. For these principles to work, cross-talk must be avoided and therefore orthogonal parts and systems are important as they are decoupled by definition. This work concerns the design and development of biological parts and tools that can enable synthetic biology in cyanobacteria. This encompasses parts necessary for the development of other systems, such as vectors and translational elements, but with a focus on transcriptional regulation. First, to enable the development and characterization of promoters in different cyanobacterial chassis, a broad-host-range BioBrick plasmid, pPMQAK1, was constructed and confirmed to function in several cyanobacterial strains. Then, ribosome binding sites, protease degradation tags and constitutive, orthogonal promoters were characterized in the model strain Synechocystis PCC 6803. These tools were then used to design LacI-regulated promoter libraries for studying DNA-looping and the behaviour of LacI-mediated loops in Synechocystis. Ultimately, this lead to the design of completely repressed LacI-regulated promoters that could be used for e.g. cyanobacterial genetic switches, and was used to design a destabilized version of the repressed promoter that could be induced to higher levels. Further, this promoter was used to implement an orthogonal transcriptional system based on T7 RNAP that was shown to drive different levels of T7 promoter transcription depending on regulation. Also, Gal4-repressed promoters for bacteria were engineered and examined in Escherichia coli as an initial step towards transferring them to cyanobacteria. Attempts were also made to implement a light-regulated one-component transcription factor based on Gal4. This work provides a background for engineering transcription and provides suggestions for how to develop the parts further.

224 citations

Journal ArticleDOI
TL;DR: Recent progresses in developing genetic tools for characterizing or manipulating cyanobacteria and algae, the applications of genetically modified strains for synthesizing renewable products such as biofuels and chemicals are critically reviewed.
Abstract: Cyanobacteria and algae are becoming increasingly attractive cell factories for producing renewable biofuels and chemicals due to their ability to capture solar energy and CO2 and their relatively simple genetic background for genetic manipulation. Increasing research efforts from the synthetic biology approach have been made in recent years to modify cyanobacteria and algae for various biotechnological applications. In the article, we critically review recent progresses in developing genetic tools for characterizing or manipulating cyanobacteria and algae, the applications of genetically modified strains for synthesizing renewable products such as biofuels and chemicals. In addition, the emergent challenges in the development and application of synthetic biology for cyanobacteria and algae are also discussed.

190 citations

Journal ArticleDOI
TL;DR: While up-regulated metabolic and antioxidative defense proteins, phytochelatin and GST work synchronously, the ars genes play a central role in detoxification and survival of Anabaena under As stress.

145 citations

Journal ArticleDOI
TL;DR: This work resulted in the construction of an improved broad-host-range replicon derived from RSF1010, which replicates in several phylogenetically distinct strains including a new experimental model strain Synechocystis sp.
Abstract: Inspired by the developments of synthetic biology and the need for improved genetic tools to exploit cyanobacteria for the production of renewable bioproducts, we developed a versatile platform for the construction of broad-host-range vector systems. This platform includes the following features: (i) an efficient assembly strategy in which modules released from 3 to 4 donor plasmids or produced by polymerase chain reaction are assembled by isothermal assembly guided by short GC-rich overlap sequences. (ii) A growing library of molecular devices categorized in three major groups: (a) replication and chromosomal integration; (b) antibiotic resistance; (c) functional modules. These modules can be assembled in different combinations to construct a variety of autonomously replicating plasmids and suicide plasmids for gene knockout and knockin. (iii) A web service, the CYANO-VECTOR assembly portal, which was built to organize the various modules, facilitate the in silico construction of plasmids, and encourage the use of this system. This work also resulted in the construction of an improved broad-host-range replicon derived from RSF1010, which replicates in several phylogenetically distinct strains including a new experimental model strain Synechocystis sp. WHSyn, and the characterization of nine antibiotic cassettes, four reporter genes, four promoters, and a ribozyme-based insulator in several diverse cyanobacterial strains.

140 citations