2 edition of Isolation, characterization and cloning of plasmids from hydrogen producing cyanobacteria found in the catalog.
Isolation, characterization and cloning of plasmids from hydrogen producing cyanobacteria
G. L. Lambert
|Statement||G.L. Lambert, N.G. Carr.|
|Contributions||Carr, N. G., Commission of the European Communities. Directorate-General for Science, Research and Development.|
The secondary metabolite hydrogen cyanide (HCN) is produced by Pseudomonas fluorescens from glycine, essentially under microaerophilic conditions. The genetic basis of HCN synthesis in P. fluorescens CHA0 was investigated. The contiguous structural genes hcnABC encoding HCN synthase were expressed from the T7 promoter in Escherichia coli, resulting in HCN production . Synechocystis produces 30 mg L-1 of RPS in 30 days (Panoff et al., ), and S. elongatus PCC can produce up to mg L-1 in 21 days (Sangar and Dugan ), values that are similar to other unicellular cyanobacteria (De Philippis and Vincenzini ) and comparably lower than the RPS production of Cyanospira capsulata, which can reach 4.
Recombinant DNA, molecules of DNA from two different species that are inserted into a host organism to produce new genetic combinations that are of value to science, medicine, agriculture, and industry. Since the focus of all genetics is the gene, the fundamental goal of laboratory geneticists is to isolate, characterize, and manipulate gh it is relatively . The production of algicidal compounds by cyanobacteria was described as early as (Harder ), and since then, many examples of allelopathy among cyanobacteria or between cyanobacteria and algae have been characterized using cultured organisms (Smith & Doan, ; Berryet al; Leãoet al, ).
Cyanobacteria are fascinating photosynthetic prokaryotes that are regarded as the ancestors of the plant chloroplast; the purveyors of oxygen and biomass for the food chain; and promising cell factories for an environmentally friendly production of chemicals. In colonizing most waters and soils of our planet, cyanobacteria are inevitably challenged by environmental stresses that . Over 2 billion years ago, with the growth of large numbers of O2-producing cyanobacteria, our planet emerged from anaerobiosis. The cyanobacteria needed only light, water, and inorganic nutrients, with CO2 as a carbon source, to replicate. Numerous genes have been identified that are required specifically for synthesis and deposition of heterocyst envelope glycolipids and .
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Introduction. Plasmids are circular or linear extrachromosomal replicons that are found in many microorganisms in the domains Bacteria, Archaea, and Eukaryota (Funnell and Phillips, ).Plasmids are transmissible by conjugation (Frost et al., ; Sota and Top, ; Frost and Koraimann, ).Smillie et al.
reported that about 14% of the full-sequenced plasmids Characterization and cloning of plasmids from hydrogen producing cyanobacteria book by: To facilitate the genetic engineering of diverse cyanobacterial strains, we have modified broad-host-range RSFbased plasmids to improve transmissibility, increase copy number, and facilitate cloning.
RSFbased plasmids replicate in diverse bacterial strains but produce low amounts of useable DNA for : Bryan Bishé, Arnaud Taton, James W. Golden. Isolation and characterization of two novel plasmids pCYM01 and pCYM02 of Cylindrospermum stagnale existence of plasmid DNA in cyanobacteria was ﬁrst observed in.
production of hydrogen. Seventy-six strains of marine photosynthetic bacteria were analyzed by agarose gel electrophoresis for plasmid DNA content. Among these strains, 12 carried two to four different plasmids with. Plasmids are important “vehicles” for the communication of genetic information between bacteria.
The exchange of plasmids transmits pathogenically and environmentally relevant traits to the host bacteria, promoting their rapid evolution and adaptation to various environments. Over the past six decades, a large number of plasmids have been identified and isolated from different Cited by: Isolation of an Allelic Series of Z-ISO Mutants in Arabidopsis and Maize.
To clone the Z-ISO gene, 12 additional mutants, 10 of which were known to be allelic to maize y9, were requested from the Maize Genetic Stock mutants all accumulated 9,15,9′-tri-cis-ζ-carotene (data not shown) as reported previously (Li et al., ).The fact that 10 mutations were allelic.
ABSTRACT CLONING, CHARACTERlZATlON AND EXPRESSION OF CARBONIC ANHYDRASE FROM THE CYANOBACTERIUM SYNECHOCYSTIS PCC Anthony Ken-Choy So. Master of Science De part ment of. Molecular hydrogen is an environmentally-clean fuel and the reversible (bi-directional) hydrogenase of the cyanobacterium Synechocystis sp.
PCC as well as the native Escherichia coli hydrogenase 3 hold great promise for hydrogen generation. These enzymes perform the simple reaction 2H+ + 2e- ↔ H2 (g). Hydrogen yields were enhanced up to fold by cloning.
PCR amplification. PCR amplification of the geoA and MIB synthase genes was carried out in 1× DynaZyme II buffer (Thermo Scientific Inc.) with μM of each primer (), μM of dNTPs, U of DynaZyme II, and 20–50 ng of genomic DNA as PCR protocol was 94 °C, 2 min, 30 cycles of 94 °C for 30s, 55 °C (geoA)/52 °C (MIB synthase) for.
production of H 2, overcoming the sensitivity of the hydrogenase enzyme to O 2, which limits the production of hydrogen in current algal systems. Approach Our goal is to construct a novel microbial hybrid taking advantage of the most desirable properties of both cyanobacteria and other bacteria, to serve as the basis.
In the prospect of engineering cyanobacteria for the biological photoproduction of hydrogen, we have studied the hydrogen production machine in the model unicellular strain Synechocystis PCC through gene deletion, and overexpression (constitutive or controlled by the growth temperature).
We demonstrate that the hydrogenase-encoding hoxEFUYH operon. Abstract. The N 2-fixing, filamentous, heterocystous cyanobacterium Nostoc PCC is capable of forming symbiotic associations with a variety of organisms, but can also grow independently from its hosts photoautotrophically as well as heterotrophically.
During the N 2-fixation process, molecular hydrogen is produced, which is metabolized by an uptake. This method may serve as a tool for the elucidation of the green alga hydrogen-related metabolism. Upon further refinement, the method may also serve in the generation of H 2 gas for the fuel and chemical industries.
The temporal sequence of events in this two-stage photosynthesis and H 2-production process is given below: (a) Green algae are grown. Vectors for cloning in cyanobacteria: construction and characterization of two recombinant plasmids capable of transformation of Escherichia coli K12 and Anacystis nidulans R2.
Mol Gen Genet. ; (2)– [Google Scholar] Meagher RB, Tait RC, Betlach M, Boyer HW. Protein expression in E. coli minicells by recombinant plasmids. Cell. SUMMARY Cyanobacteria may possess several enzymes that are directly involved in dihydrogen metabolism: nitrogenase(s) catalyzing the production of hydrogen concomitantly with the reduction of dinitrogen to ammonia, an uptake hydrogenase (encoded by hupSL) catalyzing the consumption of hydrogen produced by the nitrogenase, and a bidirectional.
Purified plasmid DNA is used in many applications from preparing vectors for cloning to generating templates for -tions. Moreover, the silica-based purification systems from Promega minimize the amount of salts and other impurities carried over during isolation, which can negatively affect downstream applications, lower yield or.
• Provide physiological and genetic characterization of the tla3 mutant, including mapping of the plasmid insert site and cloning of the gene affected in the tla3 mutation. Technical Barriers This project addresses the following technical barriers from the Biological Hydrogen Production.
Abstract. Synechococcus sp. PCC is an ideal model cyanobacterium for functional genomics and biotechnological applications through metabolic engineering. A gene expression system that takes advantage of its multiple, endogenous plasmids has been constructed in this cyanobacterium. Recombinant DNA and Gene Cloning 1.
Cloning and Expression Vectors Recombinant DNA and Gene Cloning 2. Chimeric DNA, Molecular Probes and Gene Libraries Polymerase Chain Reaction (PCR) and Gene Amplification Isolation, Sequencing and Synthesis of Genes Proteins: Separation, Purification and Identification Immunotechnology 1.
Abstract. Photosystem I is a reaction center associated with oxygenic the monomeric reaction centers in green and purple bacteria, PSI forms trimeric complexes in most cyanobacteria with a 3-fold rotational symmetry that is primarily stabilized via adjacent PsaL subunits; however, in plants/algae, PSI is monomeric.
In this study, we discovered a tetrameric. The ability of some photosynthetic microorganisms, particularly cyanobacteria and microalgae, to produce hydrogen (H2) is a promising alternative for renewable, clean-energy production.
However, the most recent, related studies point out that much improvement is needed for sustainable cyanobacterial-based H2 production to become economically viable. .The remaining plasmids were derived from pJAM and were used for DNA sequence analysis of a kb region that included the pdc gene.
Plasmid pJAM was used to produce the Z. palmae PDC protein in recombinant E. coli. The arrowhead indicates the direction of pdc gene transcription.The isolation and characterization of the lycopene ε-cyclase gene from the green microalga Chlorella (Chromochloris) zofingiensis (Czlcy-e) was performed.
This gene is involved in the formation of the carotenoids α-carotene and lutein. Czlcy-e gene encoded a polypeptide of amino acids.
A single copy of Czlcy-e was found in C. zofingiensis.