13 results about "Chlamydomonas" patented technology

The invention discloses an application of an overexpressed CrOASTL1 gene in improving the soluble protein content of leguminous forage, and belongs to the technical field of biology. 1, and the CDS sequence of the CrOASTL1 gene is as shown in SEQ ID NO. 2. The method comprises the following steps: cloning a CrOASTL1 gene segment, constructing a plant overexpression vector, carrying out plant transgenosis operation, and introducing into leguminous forage plant cells to obtain a leguminous forage transgenosis line with the overexpressed CrOASTL1 gene. The CrOASTL1 gene is over-expressed in medicago sativa, and the soluble protein content of leaves of the medicago sativa is remarkably increased by about 19%. The CrOASTL1 gene of chlamydomonas reinhardtii is applied to quality improvement of leguminous forage for the first time, the technical obstacle that the genetic relationship of species is relatively far is overcome, a brand new gene resource and an efficient technical approach are provided for cultivating a new variety of leguminous forage with high protein and high nutritional value, and the method has important significance in promoting sustainable development of grass and animal husbandry.

The application discloses a kind of biological hybrid micro machine population oral delivery system of imitating Helicobacter pylori survival mechanism, belong to oral stomach medicine delivery technical field.The system is by active power module (flagellum driven chlamydomonas reinhardtii), environmental regulation module (acid urease modified on algal surface) and multifunctional load module (macrophage membrane wrapped drug-loaded nanoparticles) synergistic assembly through polydopamine (PDA) spatial conformation control layer is formed.Active power module utilizes self-movement ability to penetrate mucus barrier and extend intracavity retention time to 12 hours or more;Environmental regulation module produces local alkalization effect by catalyzing urea hydrolysis, provides chemical shield for system in extremely acid environment of pH 1.0-3.0, protects active power module from gastric acid inactivation;Multifunctional load module is accurately delivered to gastric epithelial cells by inflammation targeting effect, such as CRISPR / Cas9 biological macromolecule, and realizes up to 31.8% in-vivo gene editing efficiency.The application provides a non-invasive, modular active delivery platform for the precise treatment of gastric diseases such as gastritis through the synergistic effect of physical penetration, metabolic reprogramming and gene intervention.

The invention discloses an eurytopic chlamydomonas reinhardtii expression vector based on a PSAD promoter and a 3 * Flag tag as well as a construction method and application of the eurytopic chlamydomonas reinhardtii expression vector. The nucleotide sequence of the eurytopic chlamydomonas reinhardtii expression vector is shown as SEQ ID No.1; the vector comprises a PSAD promoter, a target gene insertion site EcoR V, a 3 * Flag tag at the 3'terminal, an rbcS2 terminator, a TUB2 promoter, a hygromycin B screening gene and an rbcS2 terminator. The construction method comprises the following steps: constructing a skeleton of a pHyg-PSAD-3Flag vector; constructing a sequence fragment fused with a 3 * Flag tag; and connecting the sequence fragment fused with the 3 * Flag tag with a plasmid skeleton fragment through T4 DNA (deoxyribonucleic acid) enzyme so as to construct the pHyg-PSAD-3Flag vector. According to the invention, the expression plasmid of the chlamydomonas pHyg-PSAD-3Flag can be used for efficiently screening algae strains capable of stably expressing target protein.

The invention belongs to a biological deiodination technology, and provides a method for removing iodine in macroalgae and application. The chlamydomonas reinhardtii has an adsorption effect on iodine-containing substances and can be used for biological deiodination of macroalgae. Specifically, the invention provides a biological adsorption deiodination technology for removing iodine contained in edible macroalgae such as sargassum fusiforme, kelp, undaria pinnatifida and porphyra haitanensis through incubation treatment by using chlamydomonas reinhardtii in water, and a specific incubation treatment process is optimized, so that a relatively high deiodination effect is achieved.

The invention discloses a chlamydomonas engineering strain capable of stably expressing human GLP-1RA as well as a construction method and application thereof, and relates to the technical field of gene engineering. Wherein the chlamydomonas engineering strain is Aib-E4-AibRS-NMT1-GLP-1RA, the chlamydomonas engineering strain is preserved in the China Center for Type Culture Collection, and the preservation number of the chlamydomonas engineering strain is CCTCC NO: M 20252233. Through a multi-module collaborative integration strategy, the chlamydomonas engineering strain capable of synchronously realizing efficient expression of human GLP-1RA, accurate doping of alpha-aminoisobutyric acid (Aib) and myristic acylation modification is constructed. The chlamydomonas engineering strain can be used for stably and efficiently synthesizing human GLP-1RA under a photoautotrophic condition. Through verification, the obtained product is completely consistent with an original drug in the aspects of amino acid sequence, Aib doping site and myristic acylation modification mode, and meets clinical medication standards.

The application provides a Chlamydomonas reinhardtii ABC transporter gene CrABCG, a coding protein and application thereof, and the gene comprises CrABCG3 or CrABCG5, and the gene sequence comprises SEQ ID NO:1 or SEQ ID NO:2. It is found by the application that the CrABCG protein is related to carbon and nitrogen metabolism of Chlamydomonas reinhardtii, and preliminary researches show that the expression of the CrABCG3 gene of Chlamydomonas reinhardtii is increased after nitrogen limitation, while the expression of the CrABCG5 gene is first decreased and then increased, and the results show that the CrABCG protein influences the transport, synthesis and accumulation of lipid substances. Therefore, the engineering algae strains with high lipid content can be obtained by screening and modification of the algae strains with high expression of the CrABCG gene.

The invention relates to the technical field of photodynamic therapy, in particular to a synthesis method of an algae integrated oxygen supply platform based on OLED photoelectric shielding material modification, and the method comprises the following steps: step 1, selecting a photoelectric material based on pentaperylene diimide (PPD), including PPD derivatives modified by PPD-Se, PPD-Te and PPD-Br and containing heavy atoms as a core photosensitizer; step 2, mixing a photoelectric material based on PPD, a PEG derivative containing PPD-Se and a responsive chemical bond, and DSPE-PEG-S-S-NHH through a coprecipitation method, and stirring for 1-6 hours at the temperature of 30-70 DEG C to form PPD type photoelectric material nano-particles with the particle size within the range of 80-120 nm, namely PPD-based NPs; step three, pretreatment and compounding of green algae: chlorella, scenedesmus and chlamydomonas are used as oxygen supply carriers, the surface of the green algae is modified through carboxymethyl chitosan CMCS, and carboxyl groups are introduced; and step 4, combining the modified PPD type photoelectric material nano particles with the microalgae through amido bonds, so as to form the microalgae-photoelectric material composite system Alga (at) PPD-based.

The invention discloses a method for producing recombinant spider silk protein based on microalgae photosynthetic expression and non-chromatographic purification, the recombinant spider silk protein and application, and belongs to the technical field of recombinant spider silk protein production. The method comprises the following steps: designing a target recombinant spider silk protein structure, taking a spider dragline silk protein repetitive unit as a core, matching with an end-side elastin structural domain, inserting an amber termination codon TAG at a specified glycine site of the repetitive unit, and introducing non-natural amino acid containing a click chemical functional group by virtue of a gene codon extension technology; the method comprises the following steps: artificially synthesizing an optimized gene expression cassette, constructing a chlamydomonas reinhardtii expression system, synchronously constructing an unnatural amino acid insertion system, co-transforming the gene expression cassette and the unnatural amino acid insertion system into chlamydomonas reinhardtii, screening positive clones, carrying out photosynthetic culture induced expression, and carrying out pH-induced reversible phase separation in combination with magnetic nanoparticle click chemical non-chromatographic purification. Finally, the high-purity recombinant spider silk protein is prepared, and a technical basis is provided for industrialization of biological materials.

The invention discloses a recombinant beta-carotene ketolase as well as a mutant and application thereof, and belongs to the technical field of biology. According to the invention, the recombinant beta-carotene ketolase of which the catalytic activity is obviously improved is obtained by carrying out structural domain recombination on the amino acid sequences from the 1st site to the 134 site in the beta-carotene ketolase HpCrtW from haematococcus pluvialis and the amino acid sequences from the 149th site to the 328th site in the beta-carotene ketolase trCrCrtW from chlamydomonas reinhardtii; furthermore, the recombinant beta-carotene ketolase is modified through semi-rational design, so that the obtained recombinant beta-carotene ketolase mutant has relatively good catalytic activity, and the yield of canthaxanthin can be remarkably increased; furthermore, the constructed recombinant genetic engineering strain containing the recombinant beta-carotene ketolase or the mutant of the recombinant beta-carotene ketolase can produce canthaxanthin with high yield by taking glucose as a substrate, and has a remarkable industrial application value.

This invention discloses a method for rapidly isolating and purifying Rubisco from microalgae. The method includes the following steps: lysing the microalgae and filtering to obtain a supernatant of a crude protein extract; combining the supernatant of the crude protein extract with anion-exchange magnetic beads; separating the magnetic beads and eluting sequentially with a gradient elution buffer containing 40-550 mM NaCl to obtain Rubisco from the microalgae. Experiments have shown that the above-mentioned rapid method for isolating and purifying Rubisco can effectively isolate and purify Rubisco from the Chlamydomonas reinhardtii CC-125 strain. It is not only simple to operate, requiring only one step of directional enrichment, but also quick, with the entire process taking ≤3 hours. More importantly, it yields Rubisco with high purity and very few contaminating proteins. Therefore, the method provided by this invention can rapidly isolate and purify Rubisco and has significant application value.

The present invention relates to a Chlamydomonas insulata-derived 15S-lipoxygenase for simultaneously producing resolvin D5 and protectin Dx. More specifically, provided are: a composition for producing resolvin D5 and protectin Dx, the composition comprising, as an active ingredient, a Chlamydomonas insulata-derived 15S-lipoxygenase or a gene encoding same; a method for producing resolvin D5 and protectin Dx, the method comprising the step of treating a substrate with the composition; a recombinant expression vector for producing resolvin D5 and protectin Dx, the recombinant expression vector comprising the 15S-lipoxygenase gene; and a transformant in which a host cell is transformed with the recombinant expression vector.

The invention discloses a water quality purification phycomycete composition, a water quality purification system and application, and belongs to the technical field of water treatment and ecological restoration. The phycomycete composition for water quality purification comprises mixed microalgae and a composite flora, the mixed microalgae comprise chlorella sorojin, chlorella vulgaris, chlorella pyrenoidosa, chlamydomonas reinhardtii and chlamydomonas plana; the composite flora comprises bread yeast, bacillus subtilis, bacillus amyloliquefaciens, bacillus licheniformis and pseudomonas putida. According to the invention, dominant algae species in an in-situ water body and external composite flora are combined as a water quality purification phycomycete composition, so that not only is the removal efficiency of nutritive salt improved, but also the increase of microalgae biomass is promoted by improving the phycomycete microbial environment and adjusting the ecological structure. The phycomycete composition disclosed by the invention can be directly used for in-situ treatment of rivers in cold regions, is directly arranged in rivers, bank zones or constructed wetlands, and overcomes the defect that a traditional process is difficult to efficiently and stably operate in treatment of rivers in cold regions.

This invention discloses a method for constructing an engineered algal strain of high-concentration sweetness-inducing protein variants from *Chlamydomonas reinhardtii* and its applications. First, the protein coding sequences of the sweetness-inducing proteins Miraculin and Neoculin are modified and optimized to obtain optimized CrMiraculin and CrNeoculin gene sequences. These sequences are then cloned into the expression vector pGM6 to construct a recombinant expression vector. This recombinant expression vector is introduced into wild-type *Chlamydomonas reinhardtii* strains, and the transformed algal strains are selected using a paromomycin resistance marker carried by the vector. The obtained engineered algal strains can stably inherit and efficiently express the target sweetness-inducing proteins. The engineered algal strains are then fermented to obtain *Chlamydomonas reinhardtii* algal powder rich in CrMiraculin and CrNeoculin sweetness-inducing proteins. Finally, after protein purification, mixing with acidic substances can induce a sweet taste, thus demonstrating significant application potential in the food, condiment, or health product fields.