84results about How to "Improve secretion capacity" patented technology

The invention provides a novel coronavirus vaccine and a preparation method and application thereof. The encoding gene of the vaccine comprises a peptide fragment sequence, a Kozak sequence, an IgG signal peptide sequence and a human IgG Fc structural domain sequence, wherein the peptide fragment sequence contains the novel coronavirus and is combined with ACE2 in an S protein receptor binding domain RBD; the Kozak sequence and the IgG signal peptide sequence are positioned at the 5 'end of the peptide fragment sequence combined with ACE2 in the RBD; and the human IgG Fc structural domain sequence is positioned at the 3' end of the peptide fragment sequence combined with ACE2 in the RBD. The novel coronavirus vaccine has the advantages that the 1168 to 1569 sites (containing the peptide fragment combined with ACE2 in the RBD) of the encoding sequence from S protein are used as the gene sequence of antigen protein, and are modified, so that the expression ability, secretion ability and immunogenicity of the antigen protein are improved; the prepared DNA vaccine, protein vaccine and mRNA vaccine are directly injected into a human body or are injected into the human body afterbeing packaged by an adjuvant to express the corresponding antigen, and the body is induced to generate immune response; and the broad application prospect is realized in the fields of COVID-19 prevention and treatment.

The invention belongs to the technical field of microbial biology, and relates to a high-throughput pichia pastoris screening method based on a droplet micro-fluidic chip. The screening method comprises the following steps: step 1, constructing pichia pastoris; step 2, constructing a pichia pastoris mutant library; step 3, carrying out single-droplet embedding and droplet culture on strains in thePichia pastoris mutant library to obtain droplets containing Pichia pastoris mutants; step 4, carrying out droplet microfluidic sorting according to the intensity of fluorescence in the droplets, andscreening out the droplets; and 5, respectively carrying out shake-flask fermentation and re-screening on the droplets to obtain a pichia pastoris strain with improved xylanase secretion capacity. The pichia pastoris strain of xylanase fused fluorescent protein is screened by adopting a droplet microfluidic technology, and the screening flux can reach 100 thousand strains per hour; by using the method, for screening of a million-level strain library, only 10 hours are needed, and the total volume of consumed fluorescent reagents is 100 microliters.

The invention belongs to the technical field of microorganisms and particularly relates to high-conversion-rate bacillus subtilis and preparation and application thereof. The problem that the bacillus subtilis A164 is low in conversion efficiency is solved through a plasmid expression method, then a linear fragment containing a comk expression cassette is integrated on an A164 genome through an induced conversion method, and resistance is eliminated by the aid of a cre/lox specific recombination system to obtain bacillus subtilis A164S which is free of resistance marker, easy to operate and high in conversion efficiency and secretion capability. Research results show that conversion efficiency of the bacillus subtilis A164S is more than 1000 times of that of the bacillus subtilis A164; the bacillus subtilis A164S can effectively take in homologous DNA, can efficiently secrete heterologous proteins, and has high application potential in recombinant expression of exogenous genes.

The invention provides a kluyveromyces marxianus with a preservation number of CGMCC No. 10621, and also provides the application of the kluyveromyces marxianus. Compared with existing kluyveromyces marxious such as ATCC8585 and ATCC26548, the kluyveromyces marxianus has higher protein secretion capacity and higher biomass, can be used for producing enzymic preparations, and also can be used for building a recombinant protein expression system.

The invention discloses a skin care composition for controlling oil and repairing a skin barrier as well as a preparation method and application of the skin care composition. The skin care composition is prepared from the following components: saccharide isomeride, rutin and hibiscus sabdariffa flower fermentation product filtrate in a ratio of (1-5): (1-5): (1-5). The composition can be used for instantly and efficiently adsorbing grease by utilizing the saccharide isomeride, reducing excessive grease secretion of sebaceous glands in a long acting manner by utilizingthe saccharide isomeride and rutin, and moistening and repairing the skin barrier and regulating moisture-oil balance by utilizing the hibiscus sabdariffa flower fermentation product filtrate, so that the problem of facial grease production caused by excessive grease secretion of sebaceous glands is improved.

The invention discloses an intercropping feed and intercropping method for grass carps, carps, crucian carps and variegated and white carps. The intercropping feed is prepared from the following raw materials in parts by weight: 900-950 parts of a basic coarse feed, 10 parts of soybean phospholipid oil, 10 parts of soybean oil, 3 parts of sodium chloride, 10-11 parts of zeolite powder, 20 parts ofcalcium dihydrogen phosphate, 3 parts of choline chloride, 1 or less of part of Gandankang, and 10 parts of a composite premix feed for fish. According to the intercropping feed and the intercroppingmethod disclosed by the invention, physiological habits and characteristics of the grass carps, the carps, the crucian carps and the variegated and white carps are sufficiently considered, the grasscarps are mainly raised, and the intercropping feed can concurrently raise other kinds. Primary breeding test proves that components in the feed formula can generate good synergistic effects, the fishdiseases caused by factors of nutrients, environment, bacteria and the like can be effectively prevented, and the status that when being drawn in the net, the carps bleed and the carps are not resistant to transport, can be notably improved. The intercropping feed has the advantages of improving the feed digestibility, promoting growth, reducing the feed coefficient by 0.18 or above and the like,the growth rate, the daily gain and the survival rate of various intercropped fish can be effectively increased, the whole yield can be increased, and the economic benefits can be increased.

The invention belongs to the technical field of immunotherapy, and particularly relates to application of a hypoxia-adjustable promoter in CAR-T, a CAR structure of the hypoxia-adjustable promoter, anexpression vector, CAR-T cells, application of the CAR-T cells, a method for improving the IFN-gamma and/or IL-2 factor secretion capacity of the CAR-T cells in a hypoxia environment, and a method for improving the CAR-T cell killing capacity in the hypoxia environment. The CAR structure contains the hypoxia-adjustable promoter, the hypoxia-adjustable promoter is formed by connecting Hif1a adjusting elements and mini promoters, and the number of repetitions of the Hif1a adjusting elements is 3-5. The CAR structure induced and started by the hypoxia microenvironment not only can be effectivelyexpressed in T lymphocytes, but also can enhance the activation of the CAR-T cells in the hypoxia environment and enhance the effectiveness and safety of the CAR-T, and can be used for targeted therapy of tumors.

The invention discloses a method for producing 1, 3 propylene glycol safely and high efficiently, i.e. knocking out the wzi gene, which causes the generation of capsules, from K.pneumoniae through a specifically improved Red recombination system to improve the security and product secreting performance of 1, 3-PD fermentation. Compared with the prior art, the method of the invention substantially raises the gene knockout efficiency of the K.pneumoniae. The K.pneumoniae having wzi capsule gene deletion has a high product concentration, great production intensity and high security.

The invention provides a mucosal adjuvant. The mucosal adjuvant is prepared by copolymerization crosslinking compounding of oligo-chitosan and lymphotactin encoding plasmids. The invention also discloses a preparation method and a use of the mucosal adjuvant. When the mucosal adjuvant and a gene vaccine composed of chitosan or oligo-chitosan are synchronously dropped into a nasal cavity for immunization, coxsackievirus specific serum antibody and systemic (spleen and lymph glands) Th1-type immune responses are induced and especially, intestinal mucosa partially-reinforced specific-secretion-type sIgA and IFNgama+Th1 responses are induced, and thus a gene vaccine used with the mucosal adjuvant is obviously superior to an exposed gene vaccine and effectively prevents coxsackievirus-caused myocarditis. The mucosal adjuvant can be used as a novel mucosal vaccination adjuvant.

The invention relates to application of stromal vascular fraction cells and mesenchymal progenitor cells to the prevention or treatment of osteoarthritis. Specifically, remarkable prevention or treatment effects on osteoarthritis are achieved by applying a medicinal composition containing the stromal vascular fraction cells and the mesenchymal progenitor cells from autologous fat to an object with requirements. The mesenchymal progenitor cells have high cytokine secretion capability and the effect of repairing damage to organisms in a body. The stromal vascular fraction cells and the mesenchymal progenitor cells have chondrogenic differentiation and bone formation capability. The invention also provides the medicinal composition containing the stromal vascular fraction cells and the mesenchymal progenitor cells and a method for preventing and treating osteoarthritis.

The invention discloses boletus speciosus Frost polysaccharide BSF-X as well as a preparation method and an application thereof. The boletus speciosus Frost polysaccharide mainly comprises beta-D-glucose and alpha-D-galactose in the ratio being 2:1, has the average molecular weight of about 141309 Da, has a main chain of (1 arrow 4)-beta-D-glucose, a side chain of arrow 1,6)-alpha-D-galactose connected on 6-O and a arrow 4)-beta-D-glucose connected on galactose 2-O of the side chain. The boletus speciosus Frost polysaccharide has higher immunoregulation activity and can significantly or very significantly promote proliferation of three immune cells including B, T and Raw264.7; BSF-X can promote proliferation of cells, promote macrophage to release immune factors IL-23 and TNF-alpha, has aquite significant inhibition function on L929 in vitro and can inhibit growth of S180 tumor in vivo.

The invention discloses a preparation method and application of a trophoblast modified by genetic engineering, and aims to research whether K562 cells can stably express IL-21, OX40L and CD48 molecules at the same time and enhance NK cell proliferation and tumor killing ability or not. K562 gene engineering cells simultaneously expressed by the three molecules are used as trophoblasts to culture NK, so that the amplification multiple, purity and killing function of the NK are improved; according to the preparation method, K562 cells can be used for simultaneously and stably expressing membrane binding interleukin-21 (mbIL-21), OX40L molecules and CD48 molecules to prepare an engineering cell strain named as K562-ZHX trophoblast cells, then the engineering cell strain is irradiated with cobalt-60 to obtain trophoblast amplified NK cells, the activity of the NK cells prepared by the method is improved by 3 times, the cell amplification can be improved by more than 300 times, a large amount of time and cost can be saved, and a practical foundation can be laid for obtaining high-purity, high-amplification-multiple and high-killing-activity NK cells.