649 results about "Signal peptide" patented technology

The invention belongs to the field of skin care products, and particularly relates to an anti-wrinkle and anti-aging composition as well as a preparation method and application thereof, and the anti-wrinkle and anti-aging composition comprises neurotransmitter inhibition peptide, signal peptide, madecassoside, inositol and glycosylglycerol. The neurotransmitter inhibition type peptide is at least three of acetyl hexapeptide-8, arginine / lysine polypeptide, dipeptide diaminobutyrylbenzylamide diacetate and acetyl octapeptide-3, and the signal type peptide is at least two of palmitoyl pentapeptide-4, decapeptide-4 and acetyl tetrapeptide-9. Compared with the prior art, all the components of the anti-wrinkle and anti-aging composition act on different anti-aging pathways and can be matched with one another from multiple aspects of oxidation resistance, inflammation resistance, moisturizing, cell viability enhancement, wrinkle reduction and the like, a more comprehensive and better anti-aging effect is provided, skin aging is comprehensively resisted, the skin is in a more young state, and the anti-wrinkle and anti-aging composition has the advantages that the anti-wrinkle and anti-aging effects are achieved. And the anti-wrinkle and anti-aging composition is small in irritation and good in stability.

The invention relates to the technical field of biology, in particular to a pseudorabies virus gD recombinant protein, a subunit vaccine and a preparation method and application thereof.The gD recombinant protein sequentially comprises a signal peptide, a connecting peptide, the 21st-350th site of the gD protein, an enzyme cutting site and a His tag from the N end to the C end, the amino acid sequence of the 21st-350th site of the gD protein is shown as SEQ ID No.4, and the amino acid sequence of the 21st-350th site of the gD protein is shown as SEQ ID No.6. The pseudorabies virus gD recombinant protein has high immunogenicity, the prepared subunit vaccine can generate a specific antibody with neutralizing activity, mice do not die after challenge, the protection rate is 100%, the protection rate after challenge of pigs is 100%, pathological sections only show slight pathological changes, and the pseudorabies virus gD recombinant protein has wide application prospects.

The invention belongs to the technical field of biological medicine, and particularly relates to a preparation method of viroid particles displaying scFv antibodies, a product of the viroid particles and application of the viroid particles. The preparation method provided by the invention comprises the following steps: constructing a specific targeting recombinant plasmid, wherein the specific targeting recombinant plasmid is obtained by connecting a vector for expressing virus capsid protein and a coding gene for expressing an scFv antibody display structure through homologous recombination. The vector for expressing the virus capsid protein comprises a coding gene of VSV-G signal peptide, a transmembrane region and an intracellular region, and is as shown in SEQ ID NO. 2. And the coding gene for expressing the scFv antibody display structure is shown as SEQ ID NO. 3-5. According to the preparation method disclosed by the invention, the virus-like particles of which the capsid is a modular component of a specific targeting immune cell are constructed based on a lentivirus system, and 50-90% of infection efficiency can be realized in the immune cell.

The invention discloses a method for promoting lactobacillus activity enhancement, which comprises the following steps: S1, amplifying a target gene corresponding to an LPxTG structural protein, and amplifying a target gene corresponding to a signal peptide; s2, obtaining a target gene connected with the LPxTG structure and the signal peptide; s3, connecting the target gene obtained in the step S2 with a plasmid gene sequence to construct a recombinant expression vector; s4, introducing the recombinant expression vector obtained in the step S3 into escherichia coli BL-21 so as to increase the number of the recombinant expression vector; s5, introducing the recombinant expression vector obtained in the step S4 into lactobacillus plantarum C8 to obtain a recombinant strain; and step S6, co-culturing lactobacillus plantarum C8 and the recombinant strain obtained in the step S5. The lactobacillus plantarum C8 has the advantages that the whole lactobacillus plantarum C8 and the recombinant strain have better survival condition and adhesiveness in gastric juice and intestinal juice.

The invention discloses a recombinant protein for detecting a para-tumor Yo antibody through a CBA method and application, and belongs to the technical field of biomedical engineering.The amino acid sequence of the recombinant protein sequentially comprises a secretory signal peptide, a CDR2 protein partial sequence, a CDR2L protein partial sequence, a transmembrane region and a fluorescent label, and the secretory signal peptide is human albumin signal peptide ALB; the transmembrane region is a CD8a hinge; and the fluorescent label is mCherry. A novel recombinant protein which can be stably over-expressed on a cell membrane of an eukaryotic cell is constructed by intercepting specific partial sequences of CDR2 protein and CDR2L protein and redesigning and fusing a fluorescent label by using a secretory signal peptide, a transmembrane sequence and a connecting peptide, and the recombinant protein retains respective core antigen regions of the CDR2 protein and the CDR2L protein, so that the specific partial sequences of the CDR2 protein and the CDR2L protein can be stably over-expressed on the cell membrane of the eukaryotic cell. The kit can effectively overcome the defects in the aspects of sensitivity and specificity, and when a CBA method is adopted for detection, the detection rate of the para-tumor Yo antibody can be remarkably increased, and the false positive rate is reduced, so that the requirements of clinical detection are better met.

The invention relates to the technical field of active peptides, in particular to an active peptide composition and application thereof in preparation of anti-aging and wrinkle-removing products. The composition provided by the invention comprises the neurotransmitter regulating peptide and the signal peptide, and the components are properly selected, so that the composition can play a better role in anti-aging and anti-wrinkle aspects. In addition, the oil-soluble wrapped anti-aging and anti-wrinkle composition prepared by the invention aims to be quickly absorbed through skin, and the utilization rate of the active polypeptide is effectively improved.

The invention relates to the technical field of gene engineering, in particular to Escherichia coli for efficient soluble expression of recombinant human collagen III and construction and application thereof. The invention constructs Escherichia coli for efficient soluble expression of recombinant human III type collagen, and the Escherichia coli can co-express the recombinant human III type collagen and molecular chaperone which are regulated and expressed by signal peptide. Experiments prove that the engineering bacterium provided by the invention has the capability of efficient soluble expression of the recombinant human III-type collagen, and the recombinant human III-type collagen provided by the invention has good stability and anti-inflammatory effect.

The invention discloses bacillus subtilis for secretory expression of collagenase and application of the bacillus subtilis. The invention provides bacillus subtilis for secretory expression of collagenase and application of the bacillus subtilis in fat cell extraction, and after optimization of an expression element promoter and a signal peptide and large-scale fermentation of a fermentation tank, the highest enzyme activity can reach 669U. ML <-1 >. After purification, adipose-derived cells are successfully extracted and separated from mouse adipose cells, and after passage, the adipose-derived cells present the form of adipose cells. The method shows that the method has a good application prospect in the aspect of fat cell extraction.

The invention discloses a high-yield delta-tocotrienol genetically engineered bacterium and application thereof, and belongs to the technical field of gene recombination. The invention discloses a genetically engineered bacterium for high yield of delta-tocotrienols, which is characterized in that yarrowia lipolytica ATCC MYA-2613 is used as an original strain, and a ku70 gene is knocked out; the gene is used for expressing and coding 4-hydroxyphenylpyruvate dioxygenase, the gene is used for coding uromelanic acid phytate transferase and the gene is used for coding tocopherol cyclase; intensifying a shikimic acid pathway; a mevalonic acid pathway is strengthened; an isopentenol path is introduced; carrying out protein fusion on the HPD and the HPT; truncating a signal peptide of a gene encoding a tocopherol cyclase and back-filling a leucine defect. The genetically engineered bacterium constructed by the invention has industrial stability, can be amplified by more than 10 times in a 2L fermentation tank level compared with a shake flask level, can produce 616.45 mg / L of delta-tocotrienol after being cultured for 240 hours, and has industrial application prospects.

The invention discloses an E-CAR-NK cell for treating systemic lupus erythematosus and application of the E-CAR-NK cell, and belongs to the technical field of genetic engineering.A CD19 and BCMA double-target E-CAR is adopted for modifying an NK cell, and the E-CAR-NK cell is obtained; the CD19 and BCMA double-target E-CAR is composed of the following modules: a CD8 signal peptide, a CD19 scFv, a Linker, a BCMA scFv, a CD8 Hinge region, an NKG2D transmembrane region, a CD3 epsilon signal transduction region, a DAP12 costimulatory region, a 2B4 costimulatory region, and a CD3 zeta signal transduction region. Compared with a common CAR-NK cell, the E-CAR-NK cell disclosed by the invention has higher antigen sensitivity and better continuous killing ability, so that the treatment effect of SLE is improved, and the E-CAR-NK cell has lower side effects and higher safety.

The present invention relates to a bispecific chimeric antigen receptor targeting CD117 and CD7 and its application, wherein the bispecific chimeric antigen receptor comprises a signal peptide, an antigen binding domain, a hinge region, a transmembrane domain and a signal transduction domain, wherein the antigen binding domain comprises a CD117 binding domain and a CD7 binding domain. The bispecific chimeric antigen receptor provided by the present invention combines targeted antibodies for CD117 and CD7 together, recognizes and binds to two different antigens at the same time, enhances the targeted elimination ability of malignant cells, and by acting on hematopoietic stem cells and immune cells at the same time, can more effectively eliminate tumor cells in the receptor, while retaining or reconstructing normal hematopoietic and immune functions.

The invention provides a saccharomyces cerevisiae engineering strain HYB05 capable of releasing astaxanthin in a controllable self-splitting manner. The saccharomyces cerevisiae engineering strain HYB05 is constructed by taking a basic strain for producing astaxanthin as a starting strain, introducing a coding sequence comprising cell wall lyase Lyc, a coding sequence of a signal peptide mutant F8A and a recombinant nucleic acid construct for expressing a regulatory element, and knocking out an SSD1 gene to relieve a translation inhibition effect. The cell wall lyase Lyc is found and disclosed by the inventor for the first time, and the amino acid sequence is shown as SEQ ID NO: 6; the amino acid sequence of the signal peptide mutant F8A is as shown in SEQ ID NO: 8. According to the engineering strain HYB05, by introducing the recombinant nucleic acid construct, the activation of the catalytic activity of cell wall lyase under specific conditions is realized, so that the cell wall of the saccharomyces cerevisiae is broken, and the astaxanthin synthesized by the saccharomyces cerevisiae is released. Compared with the prior art, the astaxanthin is produced through fermentation of the strain, the extraction steps are simplified, extraction of the astaxanthin in the yeast is achieved to the maximum extent, the yield of the astaxanthin is remarkably increased, and the technical effect is remarkable.

The present invention relates to a viral gene therapy vector for removing hepatitis B viruses, a preparation method therefor, and the use thereof, aiming to solve the problem in which existing viral gene therapy vectors can only inhibit the replication of hepatitis B viruses, but cannot remove virus cccDNA. The vector comprises a promoter, an intron, an enhancer, a nuclear localization signal (NLS) functional domain encoding gene of PGLYRP2 protein, an HBV DNA binding domain encoding gene of the PGLYRP2 protein, a secretory signal peptide functional domain encoding gene of the PGLYRP2 protein, an HBV nucleocapsid-binding functional domain encoding gene of the PGLYRP2 protein, and an IRES fragment. The viral gene therapy vector can remarkably promote the removal of HBVs in the liver of mice and is used for treatment of hepatitis B virus infection.

The present invention discloses an improved polypeptide tumor vaccine composition and application, specifically relating to the field of polypeptide tumor vaccines, the vaccine comprises a nucleic acid sequence encoding a plurality of mutant tumor-associated antigens, the nucleic acid sequence including but not limited to: a nucleic acid sequence encoding a mutant WT1 antigen, selected from the amino acid sequence of SEQ ID NO: 1 or a variant thereof with at least 90% identity; a nucleic acid sequence encoding a mutant MUC1 antigen, selected from the amino acid sequence of SEQ ID NO: 2 or a variant thereof with at least 90% identity; a nucleic acid sequence encoding a mutant PD-1 antigen, selected from the amino acid sequence of SEQ ID NO: 3 or a variant thereof with at least 90% identity; and the vaccine composition further comprises one or more modified elements that enhance antigen expression or immune response, including a signal peptide, an adjuvant coding sequence or an immunostimulatory sequence. The vaccine composition of the present invention is intended to enhance the immune response to a plurality of tumor antigens and improve the effectiveness of tumor treatment.

The present invention belongs to the technical field of gene therapy. Provided are a chimeric antigen receptor nanocomposite and a preparation method therefor and the use thereof in the preparation of an anti-aging drug. The provided chimeric antigen receptor nanocomposite comprises a chimeric antigen receptor DNA plasmid specifically recognizing senescent cells, a polymer coupled with a signal peptide, a free polymer and an antibody polymer targeting T cells, wherein the chimeric antigen receptor DNA plasmid, the polymer coupled with a signal peptide, the free polymer and the antibody polymer targeting T cells are sequentially mixed and incubated to form the chimeric antigen receptor nanocomposite. The chimeric antigen receptor nanocomposite can rapidly generate senescent cell-marked chimeric antigen T cells in vivo. The engineered T cells have the ability to specifically eliminate senescent cells in vivo, and can rapidly and durably eliminate senescent cells in vivo, which is irreplaceable and has wide application prospects.

The invention relates to the technical field of antibacterial peptides, and discloses a preparation method and application of a multi-segment fused antibacterial peptide fusion protein. The antibacterial peptide fusion protein is obtained by introducing OST signal peptide and collagen and optimizing the connection sequence of the OST signal peptide, the collagen, bovine lactoferrin peptide LFcinB, bovine lactoferrin N-leaf small peptide LFcin and bovine lactoferrin N-leaf derived peptide Lf. The antibacterial peptide fusion protein can simultaneously contain three segments of bovine lactoferrin active peptides, namely LFcinB, LFcin and Lf, and the yield of the fusion peptide is maximized. Experiments prove that the fermentation yield of the fusion protein comprising the OST signal peptide, the collagen, the bovine lactoferrin peptide LFcinB, the bovine lactoferrin N-leaf small peptide LFcin and the bovine lactoferrin N-leaf derived peptide Lf which are sequentially connected from the N terminal to the C terminal can reach 600mg / L.

The invention discloses a pichia pastoris high-secretion strain screening system and a use method thereof. According to the system, pichia pastoris is used as a host, and the core structure of the system is as follows: an AOX1 promoter, an alpha-factor signal peptide, a target protein-P2A peptide, an alpha-factor signal peptide, a Gamilus fluorescent protein-G4S connecting peptide-Gcw21 anchoring sequence-AOX1 terminator. The use method comprises the steps of vector construction, conversion of enzyme digestion recovery products, clone strain selection and activation, fermentation, inducible expression and screening, and product collection to finally obtain the strain with high yield of target protein. Through integration of bicistronic design, P2A peptide self-cleavage and cell surface display technologies, efficient screening of target protein high-secretion type strains is realized.

The invention belongs to the technical field of bioengineering, and particularly relates to an IL-12-CD137 chimeric protein, a function-enhanced engineered NK cell, and preparation and application thereof. The IL-12-CD137 chimeric protein comprises a signal peptide, an IL-12a protein molecule, a CD137 transmembrane structural domain and a CD137 intracellular structural domain, and has an amino acid sequence as shown in SEQ ID NO: 1. According to the function-enhanced engineered NK cell, the IL-12-CD137 chimeric protein sequence is introduced into an original NK cell, the function-enhanced engineered NK cell expressing Chime-IL12-CD137 is obtained, the amplification capacity and killing capacity of the NK cell can be enhanced, and the function-enhanced engineered NK cell is applied to treatment of tumor and virus-infected diseases of companion animals for the first time.

The invention discloses a recombinant I-type collagen, a preparation method and application, and belongs to the technical field of binding tissue peptides, the recombinant I-type collagen is formed by sequentially connecting the following modules in series from the N terminal to the C terminal: a signal peptide, a truncated I-type collagen and a His tag; the nucleotide sequence of the truncated I type collagen is as shown in any one of SEQ ID NO.3, SEQ ID NO.4, SEQ ID NO.5 and SEQ ID NO.6; the nucleotide sequence of the truncated I type collagen is as shown in any one of SEQ ID NO.3, SEQ ID NO.4, SEQ ID NO.5 and SEQ ID NO.6; according to the invention, a mammalian expression system and a 293T cell line are used for expressing recombinant I-type collagen, so that collagen with natural biological activity is obtained, and the proliferation efficiency and repair capacity of the collagen for promoting HSF cells can be improved.

The invention discloses an influenza C virus HEF protein polyclonal antibody preparation method, which comprises: carrying out signal peptide prediction, antigen epitope screening and hydrophobicity analysis through bioinformatics, constructing a recombinant prokaryotic expression vector pET32a-HEF by using a seamless cloning technology, and efficiently expressing the HEF protein in Escherichia coli to obtain the influenza C virus HEF protein polyclonal antibody. The inclusion body protein yield is improved by optimizing conditions; and purifying by nickel column affinity chromatography to obtain high-purity recombinant protein, emulsifying the high-purity recombinant protein and a Freund's adjuvant, immunizing a Japanese white rabbit, immunizing for three times, and collecting high-titer serum. The method breaks through detection limitation caused by antigen tag concealment in a eukaryotic expression system, the obtained polyclonal antibody is high in specificity, the eukaryotic expression HEF protein can be accurately recognized, and a key tool is provided for ICV virus-like particle identification, vaccine research and development and diagnosis technologies. Compared with a traditional method, the scheme is simple and convenient to operate, low in cost and high in antibody titer, and has remarkable application value.

The invention provides functional identification and application of a novel cell virulence factor. Based on the research on the novel virulence factor of the cell, the invention also provides a polypeptide delivery system and application thereof. Wherein the polypeptide delivery system comprises a protein complex and a signal peptide, the signal peptide comprises at least 20 amino acids at the N-terminal of PagT1 protein or homologous protein thereof, and the signal peptide is used for guiding the polypeptide to be loaded into the protein complex and guiding the polypeptide to be positioned to the nucleus of eukaryotic cells. The research of the invention provides an important basis for constructing a safe and efficient novel tumor treatment system, and provides an innovative solution for treating serious diseases such as cancers.

The invention provides low-temperature alginate lyase and application thereof, namely alginate lyase with low-temperature catalytic activity, which is separated from marine-derived cold monad bacteria, and the amino acid sequence of the alginate lyase is SEQ ID NO: 1; the amino acid sequence of the alginate lyase after the signal peptide is removed is SEQ ID NO: 3. The algin lyase Alg3889 provided by the invention has the optimal reaction temperature of about 10-16 DEG C, has good temperature stability, and still keeps more than 70% of enzyme activity after being incubated at 0-70 DEG C for 1 hour; the optimum pH value of the strain is 8.0, after the strain is incubated in a buffer system with the pH value of 3.0-10.6 for 12 h, about 90% of the highest enzyme activity can be kept, and the strain has high acid resistance and alkali resistance. The degradation product of the alginate oligosaccharide is alginate oligosaccharide with the polymerization degree of 2-6, and the alginate oligosaccharide has high application value.

The invention discloses a variable region amino acid sequence of a monoclonal antibody of a West Nile virus non-structural protein NS1 and application of the variable region amino acid sequence, and belongs to the technical field of medicines. According to the invention, West Nile virus non-structural protein NS1 expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the West Nile virus non-structural protein NS1 are screened from rabbit spleen cells through flow sorting, and a signal peptide and a variable region gene fragment of an antibody are cloned through reverse transcription-polymerase chain reaction; according to the present invention, the non-structural protein NS1 of flaviviridae flaviviridae virus is taken as a template, and is connected with a constant region gene to an expression vector, and after mammalian cell expression and purification, the monoclonal antibody which has high affinity and is not combined with the non-structural protein NS1 of other eight viruses of flaviviridae flaviviridae virus is obtained through enzyme-linked immunosorbent assay; the monoclonal antibody has application value in diagnosis and prevention and treatment of West Nile virus infection.

The composition is prepared from group B meningococcus NHBA recombinant protein, fHbp (V1-V1) recombinant protein and fHbp (V2-V3) recombinant protein obtained through artificial modification, or three corresponding recombinant proteins obtained by further adding P4 signal peptide on the basis of the group B meningococcus NHBA recombinant protein, the fHbp (V1-V1) recombinant protein and the fHbp (V2-V3) recombinant protein, and animal experiments prove that the composition has good immunogenicity and can be used for preparing the immunopotentiator. The compound can be used for preparing vaccines for preventing group B meningococcus.

The invention belongs to the technical field of biology, and particularly relates to recombinant human collagen III and an expression method of the recombinant human collagen III. The amino acid sequence of the recombinant human collagen III is shown as SEQ ID No.1; the amino acid sequence of the recombinant human III type collagen comprises a plant secretion signal peptide Apo, a human III type collagen Gln24-Leu 1466 segment and a 6His tag. Besides, tobacco is used as a carrier, an agrobacterium tumefaciens-mediated instantaneous conversion technology is applied, efficient and stable expression of the recombinant human-derived III-type collagen is achieved, the ethical and safety problems of a traditional collagen source are effectively solved, and a new way is established for production of recombinant protein. The innovative achievement has wide application prospect and important scientific value, and is expected to promote sustainable development and progress in the field of biotechnology.

The invention discloses a signal peptide SEQ ID NO: 1-3 suitable for yeast, the coding gene of the signal peptide is expressed in the yeast after being fused with a foreign protein gene, the expression level of the foreign protein can be improved, and the signal peptide has popularization and application prospects.

A system for delivering a payload nucleic acid into target cells of a subject and production of a payload (or payloads) encoded by the payload nucleic acid in the cells. The system includes a Bifidobacterium sp. bacterium comprising a plasmid and a transporter nucleic acid, the transporter nucleic acid configured for expression in the bacterium. The transporter nucleic acid encodes a transporter polypeptide comprising, in an amino-terminal to carboxy-terminal order, a bacterial secretion signal peptide, a DNA-binding domain to bind the plasmid, and a cell penetrating peptide. The transporter polypeptide complexes with the plasmid and transports the plasmid from the bacterium into the target cells. The plasmid encodes one or more payloads (protein and / or ribonucleic acid) for production in the target cells. The target cells may be colonic cells. When the payload(s) include an antigen, the system may be a DNA vaccine.

Described herein are engineered cytokine receptor switches that can include a signal peptide, an extracellular activator binding domain, a hinge, a transmembrane domain, and / or an intracellular signaling domain. Binding of an activator to the activator binding domain can activate cytokine signaling through the intracellular signaling domain. These cytokine receptor switches can be expressed in immune cells, sometimes in combination with a chimeric antigen receptor (CAR), to increase immune cell persistence by promoting adoption of memory-like phenotypes. Also described herein are methods of using engineered cytokine receptors in immune cell therapies, such as CAR T-cell therapy, to improve patient outcomes and prevent disease relapse.

The invention discloses recombinant bacillus subtilis capable of efficiently expressing a plastic degrading enzyme LCCICCG as well as a construction method and application of the recombinant bacillus subtilis. The recombinant bacillus subtilis is used for overexpressing the plastic degrading enzyme LCCICCG and signal peptide. By constructing plasmids of different signal peptides and promoters, the biomass of the recombinant bacillus subtilis and the protein expression quantity of the LCCICCG in the fermentation culture process are improved, and the industrial production cost of the LCCICCG is reduced.

The invention relates to pichia pastoris for producing brazitame protein as well as a construction method and application of the pichia pastoris, and belongs to the technical field of microorganisms. ExpL SP is used as a signal peptide in pichia pastoris, heterologous expression of Brazilian sweet protein is carried out through an AOX1 promoter, the ExpL SP and a Pro region of an alpha mating factor are further fused and transformed to obtain a new signal peptide to enhance the secretion efficiency, and the new signal peptide and a ribosomal protein synthesis factor Bcy1 are co-expressed to improve the protein synthesis capacity. The constructed pichia pastoris can be used for efficiently secreting the Brazilian sweet protein, the problems that in the prior art, when microorganisms are used for fermenting the Brazilian sweet protein, the Brazilian sweet protein is mistakenly folded, the sweet taste is reduced or lost are solved, the shake flask yield is stabilized at 450 mg / L, the fermentation tank yield reaches 5.215 g / L, and a high-cost-performance scheme is provided for industrial production of the Brazilian sweet protein.