34 results about "Chlamydomonas" patented technology

The invention discloses application of a CMIET protein and a coding gene thereof in improving manganese resistance of plants. The invention belongs to the technical field of biology, and particularly relates to application of CMIET protein and a coding gene thereof to improvement of manganese resistance of plants. The CMIET protein can regulate and control biological manganese resistance, and excessive manganese in Chlamydomonas chloroplast can be discharged to cytoplasm by regulating and controlling expression of the protein CMIET or regulating and controlling gene or regulating and controlling activity or content of the protein CMIET, so that balance of manganese ion concentration in the chloroplast is maintained, and the chloroplast is prevented from being affected by manganese toxicity; the normal photosynthesis capability of the cells is maintained, and the normal photosynthesis is ensured.

The invention provides an SNP (Single Nucleotide Polymorphism) site related to the temperature-resistant character of chlamys farreri and an application of the SNP site in breeding, namely the SNP site which is located on a No.12 chromosome of the chlamys farreri and is related to the temperature-resistant character, the temperature resistance of the chlamys farreri is evaluated based on the typing condition of the SNP site, and a molecular marker is provided for breeding of a new temperature-resistant variety of the chlamys farreri. The SNP site is located at the 554th site of the molecular marker with the sequence of SEQ ID NO: 1, and is T / G. According to the invention, the SNP sites related to the temperature resistance character of the chlamys farreri are obtained through screening, and a method for screening the chlamys farreri is established. The method is more suitable for whole genome selective breeding work, parents are rapidly screened according to the typing condition of the SNP, individuals with the genotype being TT at the site can be preferentially selected in production to serve as parents for breeding temperature-resistant scallops for breeding, and GG type individuals are prevented from being brought into the parents.

The invention discloses a general expression plasmid pHyg-HSRB-intron-3HA vector for Chlamydomonas reinhardtii as well as a construction method and application of the general expression plasmid pHyg-HSRB-intron-3HA vector. A nucleotide sequence of the general expression plasmid The vector comprises an HSRB promoter, an RBCS2intron, a target gene insertion site EcoRV, a 3 * HA tag at the 3'terminal, an rbcS2 terminator, a TUB2 promoter, a hygromycin B (Hyg +) screening gene and an rbcS2 terminator. The construction method comprises the following steps: constructing a skeleton of an expression plasmid pHyg-HSRB-intron-3HA vector; and constructing a combined fragment of an HSRB promoter and RBCS2intron, and carrying out homologous recombination on the combined fragment and a plasmid skeleton structure fragment to construct a vector for expressing plasmid pHyg-HSRB-intron-3HA. The method can improve the acquisition rate of positive transformants and the stable high expression of target genes.

The invention discloses a treatment method of fermentation wastewater. The treatment method of the fermentation wastewater comprises the following steps: adding filamentous fungi and green algae into fermentation wastewater generated after fermentation treatment of kitchen garbage, carrying out fermentation culture, then carrying out solid-liquid separation, and extracting grease from the separated solid. The filamentous fungi comprise trichoderma harzianum, and the green algae comprise one or a combination of more selected from chlorella vulgaris, chlamydomonas, haematococcus pluvialis, hylothrix, scenedesmus obliquus and chaetoclada. According to the treatment method of the fermentation wastewater, grease can be produced while the fermentation wastewater is treated, and the treatment cost of the fermentation wastewater is reduced.

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.

Provided is a composition comprising Chlamydomonas reinhardtii or an extract of Chlamydomonas reinhardtii, the Chlamydomonas reinhardtii or the extract of Chlamydomonas reinhardtii comprising an agonist of an opioid receptor.

Provided is a composition comprising Chlamydomonas reinhardtii or an extract of Chlamydomonas reinhardtii, the Chlamydomonas reinhardtii or the extract of Chlamydomonas reinhardtii comprising an agonist of a progesterone receptor.

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 present invention discloses a rabbit polyclonal antibody against Chlamydomonas reinhardtii NPHP4 protein, its preparation method and application. The amino acid sequence of the NPHP4 protein is shown in SEQ ID NO.1. Preparation method: Analysis and design of the antigen epitope of Chlamydomonas reinhardtii NPHP4 protein, and selection of the 859-1058aa amino acid fragment to construct a prokaryotic cell recombinant expression vector; transferring the recombinant expression vector into prokaryotic expression cells to obtain a recombinant expression strain and inducing the expression of the antigen protein; using the purified antigen protein as an immunogen to immunize rabbits to obtain a rabbit polyclonal antiserum against the NPHP4 protein, and further purifying it; the purified polyclonal antibody can be used to verify the labeling of the ciliary transition zone of Chlamydomonas reinhardtii by the IF method, proving that it can be used for specific fluorescence labeling of the ciliary transition zone; it can also be used for qualitative and quantitative analysis of the Chlamydomonas reinhardtii NPHP4 protein by the immunoblotting WB method.

The invention discloses a method for efficiently synthesizing recombinant human lactoferrin, which comprises the following steps: transforming a target gene into Rhin microalgae at least twice to construct microalgae engineering bacteria, the target gene being a gene capable of expressing human lactoferrin; the method comprises the following steps: inoculating microalgae engineering bacteria into a liquid culture medium, carrying out illumination culture at the temperature of 20-30 DEG C, harvesting algae liquid, and obtaining the recombinant human lactoferrin from the algae liquid. The human lactoferrin HLF gene is transformed at least twice in the chlamydomonas reinhardtii, a human lactoferrin synthesis route is introduced, an engineering algal strain with high expression quantity is constructed, a culture medium formula is adjusted on the basis, culture is performed under a 5L fermentation amplification condition, finally, the maximum yield of the human lactoferrin is increased by about 7 times compared with that before culture condition optimization, and the yield of the human lactoferrin HLF gene is increased by about 7 times compared with that before culture condition optimization. And 1.1% of the dry weight of the cells is achieved.

To provide a modified channel rhodopsin having high ion permeability (photoreactivity).SOLUTION: A third extracellular domain counting from the N-terminal side of three extracellular domains possessed by channel rhodopsin derived from volvox is replaced with an extracellular domain corresponding to channel rhodopsin-2 derived from Chlamydomonas reinhardtii.SELECTED DRAWING: Figure 2

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 preparation method of chlamydomonas reinhardtii hydroxyproline-rich glycoprotein, which belongs to the technical field of biological medicine, and comprises the following steps: soaking fresh and alive chlamydomonas reinhardtii cells in an alcoholic solution, re-suspending the cells in a water phase, adding grinding beads, oscillating, centrifugally collecting a white precipitation layer, dissolving the white precipitation layer in a solubilizing solvent, filtering, and drying to obtain the chlamydomonas reinhardtii hydroxyproline-rich glycoprotein. And purifying by adopting ultrafiltration or gel column chromatography to obtain the hydroxyproline-rich glycoprotein of the chlamydomonas reinhardtii. The hydroxyproline content in the prepared hydroxyproline-rich glycoprotein of the chlamydomonas reinhardtii can reach 188 amino acid residues per 1000 amino acid residues, and glycosylation modification is mostly single galactoside, single arabinoside or double arabinoside. The invention provides a microalgae-derived collagen analogue which can be applied to the fields of foods, health foods, cosmetics, medicines and the like.

The present invention discloses a method for efficiently synthesizing recombinant human lactoferrin, comprising: transforming a target gene at least twice into Chlamydomonas reinhardtii microalgae to construct a microalgae engineering bacterium, wherein the target gene is a gene capable of expressing human lactoferrin; inoculating the microalgae engineering bacterium into a liquid culture medium and culturing under light at a temperature of 20 to 30°C, harvesting the algae liquid, and obtaining recombinant human lactoferrin from the algae liquid. The present invention transforms the human lactoferrin HLF gene at least twice in Chlamydomonas reinhardtii, introduces a human lactoferrin synthesis pathway, constructs an engineered algae strain with a high expression level, adjusts the culture medium formula on this basis, and cultivates under 5L fermentation amplification conditions, and finally achieves a maximum yield of human lactoferrin that is about 7 times higher than before the optimization of the culture conditions, reaching 1.1% of the cell dry weight.

The invention belongs to the technical field of gene engineering, and particularly relates to a cre-FTO gene for improving the carbon sequestration efficiency of chlamydomonas reinhardtii, a related biological material and application. The cre-FTO gene disclosed by the invention is an RNA (Ribonucleic Acid) demethylase gene, and the nucleotide sequence of the cre-FTO gene is shown as SEQ ID NO: 1. The recombinant expression vector pDb124-cre-FTO containing the cre-FTO gene is transformed into the chlamydomonas reinhardtii, so that the photosynthetic efficiency, the carbon sequestration efficiency and the biomass of wild type chlamydomonas reinhardtii can be improved. The cre-FTO gene and related biological materials provided by the invention are helpful for improving the photosynthetic efficiency, starch conversion rate, pyruvic acid conversion rate and other carbon sequestration efficiencies of chlamydomonas reinhardtii, and have a more promising application value in improving the photosynthetic efficiency and the carbon sequestration efficiency.

Provided is a composition comprising Chlamydomonas reinhardtii or an extract of Chlamydomonas reinhardtii, the Chlamydomonas reinhardtii or the extract of Chlamydomonas reinhardtii comprising an antagonist of a histamine receptor.

The invention discloses a rabbit polyclonal antibody of chlamydomonas reinhardtii NPHP4 protein as well as a preparation method and application of the rabbit polyclonal antibody. The amino acid sequence of the NPHP4 protein is shown as SEQ ID NO. 1. The preparation method comprises the following steps: analyzing and designing a chlamydomonas reinhardtii NPHP4 protein antigen epitope, and selecting 859-1058aa amino acid fragments to construct a prokaryotic cell recombinant expression vector; transferring the recombinant expression vector into a prokaryotic expression cell to obtain a recombinant expression strain, and inducing antigen protein expression; carrying out rabbit immunization by taking the purified antigen protein as an immunogen to obtain NPHP4 protein rabbit polyclonal antiserum, and further carrying out purification treatment on the NPHP4 protein rabbit polyclonal antiserum; the purified polyclonal antibody can be used for labeling verification of a cilia transition region of Chlamydomonas reinhardtii by an IF method, and the result proves that the polyclonal antibody can be used for specific fluorescence labeling of the cilia transition region; and the chlamydomonas reinhardtii NPHP4 protein can also be qualitatively and quantitatively analyzed by an immunoblotting WB method.

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 relates to the technical field of food processing, and in particular to a method for preparing a low-fat, low-calorie, algae-free protein drink, comprising: S1, the algae raw material is a chlorophyll-deficient high-protein microalgae, the algae cell protein content is ≥35%, the oil content is ≤15%, and the starch content is ≤30%; S2, water is added to adjust the dry weight of the microalgae cells to 50-150 g / L, and a high-pressure homogenizer is used to crush the cells to obtain a cell slurry; if the microalgae raw material is a slender euglensis without a cell wall or a Chlamydomonas reinhardtii with a cell wall as a polysaccharide protein, the cells are homogenized 1-2 times; if the microalgae raw material is a microalgae with a cell wall and the main component of the cell wall is cellulose, high-pressure homogenization is required until the particles below 1 μm in the cell slurry account for more than 90% of the total number of particles, and the particles below 300 nm account for more than 50% of the total number of particles; S3, one or more of cellulase, protease, amylase and saccharifying enzyme are added according to the type of microalgae for enzymolysis, and a pH regulator is added after the enzymolysis is completed; S4, homogenization is performed again; S5 sterilization and filling. The microalgae protein beverage of the present invention can better meet the needs of people who are allergic to milk or lactose intolerant or who pursue low calorie intake.

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.

An object of the present invention is to provide a modified channelrhodopsin having high ion permeability (photoreactivity). The solution is to substitute the third extracellular domain counted from the N-terminal side of three extracellular domains included in a Volvox carteri-derived channelrhodopsin by a corresponding extracellular domain of a Chlamydomonas reinhardtii-derived channelrhodopsin-2.

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.

The utility model discloses a micro-fluidic chip for observing chlamydomonas, which belongs to the field of biological experiment equipment, the micro-fluidic chip for observing chlamydomonas comprises a substrate, a cover plate, a first stop valve and a second stop valve, the upper surface of the substrate is provided with an accommodating cavity, and the substrate is provided with a first lead-in channel, a second lead-in channel and a lead-out channel which are communicated with the accommodating cavity. A cover plate covers the upper surface of the base plate and covers the containing cavity, the first guide-in channel, the second guide-in channel and the guide-out channel. And the first stop valve controls the first lead-in channel to be opened and closed. And the second stop valve controls the connection and disconnection of the second lead-in channel. According to the micro-fluidic chip for observing the chlamydomonas provided by the utility model, one of the first lead-in channel and the second lead-in channel can be closed in the process of observing the chlamydomonas, so that a final observation result is prevented from being influenced by uncontrolled mixing of a chlamydomonas culture solution or medicine in the process of observing the chlamydomonas.

Provided is a composition comprising Chlamydomonas reinhardtii or an extract of Chlamydomonas reinhardtii, the Chlamydomonas reinhardtii or the extract of Chlamydomonas reinhardtii comprising an antagonist of a cysteinyl leukotriene receptor.