151results about How to "Efficient secretion" patented technology

The present invention is directed to a method and apparatus for electrical stimulation of the pancreatico-biliary system. Electrode sets are placed in the pancreatico-biliary system in an arrangement that induce contractions or relaxation of the portion or whole of the pancreatico-biliary system by electrical stimulation of the surrounding tissue, muscles and nerves. The electrical stimulus is applied for periods of varying duration and varying frequency so as to produce the desired therapeutic effect, including inhibiting fat digestion or fat absorption by a patient and inducing satiety in the patient.

The present invention provides delivery vectors for transferring a nucleic acid sequence to a cell in vitro, ex vivo or in vivo. The delivery vector comprises a segment encoding a secretory signal peptide. In embodiments of the invention, the delivery vector is an adeno-associated virus (AAV) vector. In other embodiments, the secretory signal peptide is a fibronectin secretory signal peptide (including variations and modifications, thereof). The delivery vectors of the invention may further comprise a heterologous nucleic acid sequence encoding a polypeptide of interest for transfer to a target cell, where the polypeptide of interest is operably associated with the secretory signal. Also disclosed are methods of transferring a nucleic acid of interest to a cell using the delivery vectors of the invention.

A DNA encoding 20K hGH is connected directly to a gene encoding Escherichia coli OppA protein secretion signal, or a modified form thereof, and a DNA encoding signal peptidase 1 to construct a recombinant plasmid, E. coli is transformed by said plasmid and cells of the resulting E. coli transformant strain are cultured for secretory production of the 20K hGH in the E. coli periplasm. This method enables efficient secretory production of 20K hGH and easy isolation and purification of 20K hGH from the periplasm fraction because the level of impure proteins in the E. coli periplasm is low.

The invention provides a CP7 strain that can produce new antibacterial polypeptides that have strong antibacterial effect to gram-positive bacteria and are small peptides with 23 amino acid residues by the determination and whose amino acid sequence is: LLLAQPTAAVRGRCQVQRYLLA A. The invention also provides the application of the new antibacterial polypeptide in the preparation of the anti gram-positive bacteria drug.

The invention discloses an in-vitro large-scale amplification method of natural killer cells. The method comprises the following steps: collecting and separating peripheral blood mononuclear cells; sorting natural killer cells; culturing the natural killer cells; and collecting the natural killer cells. The method utilizing immunomagnetic beads to sort the natural killer cells has the advantages of simplicity in establishing and operation, and high comprehensive efficiency; and the effect of the method adopting in-vitro amplification of the immunomagnetic bead shorted NK cells is 2-3 times the effect of present amplification methods, so the method disclosed in the invention has great application values in the field of in-vitro large-scale amplification.

The present invention provides a method for efficiently producing an industrially useful protein in coryneform bacteria, and more particularly, a method for efficiently producing a protein for which secretion was difficult using conventional protein secretion pathways. In particular, the present invention provides a method for efficiently producing heterologous proteins comprising culturing coryneform bacteria containing an genetic construction containing a promoter sequence which functions in coryneform bacteria, a nucleic acid sequence encoding a Tat system-dependent signal peptide region, and a nucleic acid sequence encoding a heterologous protein, in the direction from 5′-end to 3′-end, and secretory producing the heterologous protein by coryneform bacteria.

This invention relates to a novel protein having a lecithin-cholesterol acyltransferase-like activity, etc. or its salt, a precursor protein of the protein or its salt, a partial peptide of the protein or its salt; a DNA coding for the protein; a recombinant vector; a transformant; a method for producing the protein, a pharmaceutical composition comprising the protein, the partial peptide or its salt; and an antibody to the protein or the partial peptide. The protein, the partial peptide or its salt, and the DNA are useful as an agent for treating or preventing arteriosclerosis, atherosclerosis, hyperlipidemia, hypercalorism, obesity or hypertriglyceridemia. The antibody can be used in assay of the protein, the partial peptide or its salt. The protein, the partial peptide or its salt is useful as a reagent for the screening for candidate medical compounds.

The present invention relates generally to the use of recombinant adeno-associated viruses (rAAV) for gene delivery and more specifically to the use of rAAV to deliver antibody genes to target cells in mammals. Administration of rAAV encoding antibodies that neutralize the HIV-1 virus is exemplified.

Provided are a grip strength detection mechanism, an exercise apparatus provided with the same, and a method for using the same. The exercise apparatus is provided with: a grip part 12 comprising a cylindrical flexible member; sealing members 13, 16 which seal both ends of the grip part 12 to form a deformable sealed space 18 which is surrounded by the grip part 12; a pressure sensor 28 which is held by the sealing member 16 and detects the pressure within the sealed space 18; and a control unit which detects, as the pressure within the grip part, a reference grip strength which is not more than the maximum grip strength detected by the pressure sensor 28 in order to display, on a display unit 7, the grip strength of a user gripping the grip part 12.

The invention discloses recombinant engineering bacteria for efficiently expressing human growth hormone (hGH), a construction method and an application and provides an escherichia coli operon for expressing recombinant hGH, an expression plasmid containing an operon sequence and engineering bacteria QJSW-SZ01 (CGMCC No:7258) used for secretory expression of the recombinant hGH and obtained by transformation of the expression plasmid containing the operon. The recombinant engineering bacteria are characterized in that a coding sequence of a signal peptide of an escherichia coli heat-stable enterotoxin is changed, and rare codons of escherichia coli in the coding sequence are mutated so as to avoid formation of a secondary structure; meanwhile, an amino acid is altered on the terminal of the coding sequence so that the coding sequence is more beneficial to guidance of secretory expression of hGH; the signal peptide, an escherichia coli alkaline phosphatsae promoter (phoA promoter) and an escherichia coli T7 terminator are combined to be used as an expression control element so that the recombinant hGH is efficiently secretory-expressed in the escherichia coli in a soluble form. The recombinant engineering bacteria lay the foundation of finally developing a low-cost hGH pharmaceutical product.

The invention discloses skin cream containing camellia seed oil. The skin cream containing camellia seed oil comprises 100 parts of water, 0.5 to 5 parts of p-hydroxyacetophenone, 1 to 8 parts of betaine, 0.1 to 0.8 parts of carbomer, 5 to 15 parts of glycerin, 5 to 15 parts of butanediol, 0.05 to 0.3 parts of sodium hyaluronate, 1 to 10 parts of nicotinamide, 0.05 to 0.5 parts of dipotassium glycyrrhizinate, 10 to 25 parts of camellia seed oil, 5 to 15 parts of mink oil, 1 to 5 parts of cetearyl alcohol and / or cocoyl glucoside, 2 to 10 parts of jojoba seed oil, 2 to 5 parts of stearate, 2 to10 parts of avocado fruit fat, 2 to 10 parts of cyclopentadimethylsiloxane, 2 to 10 parts of squalane, 2 to 10 parts of polydimethylsiloxane, 2 to 10 parts of isodecyl isononanoate, 3 to 10 parts of triethylhexanoin, 1 to 10 parts of tocopherol, 0.1 to 0.8 parts of triethanolamine, and 0.5 to 5 parts of 1, 2-hexanediol. The invention discloses a preparation method of the skin cream containing camellia seed oil. The preparation method utilizes scientific combination of the raw materials and realizes moisturizing, wrinkle resistance, aging resistance, sunscreen and skin repair. The product is refreshing and non-greasy and is suitable for different types of skin.

Modified viral particles wherein the viral particles, typically adenoviral particles, are modified by glycosylation and the use of the modified viral particles to deliver heterologous nucleic acid to cells. Also disclosed are pharmaceutical compositions comprising the same and method of treatment using the same.

A high temperature alpha-amylase and a high efficiency preparation method of the mutant thereof belongs to the fermentation engineering field. The invention provides a method for high efficiently secreting expressing a high temperature alpha-amylase and mutant thereof with alpha-amylase activity through a bacillus, especially a bacillus licheniformis mutant strain as a host cell. The synthesis and secretion level of the high temperature alpha-amylase or mutant thereof of the recombinant strain of the invention reaches 18-25mg / mL during fed-batch fermentation and is 3-4.5 times of the original strain high temperature alpha-amylase synthesis level. The high temperature alpha-amylase of the invention is useful for reducing the fermentation manufacture cost of the industrial enzyme, simplifying the fermentation manufacture process and reducing fermentation industrial environment pressure.

Provided are means for evaluating and identifying putative substrates of the twin arginine translocation (Tat) secretory pathway in Streptomyces and other bacterial species. Also provided, therefore, are simple ways to express, secrete and purify correctly folded heterologous proteins on a large scale using host microorganisms, such as, Streptomyces and the Tat pathway therein. Many of the thus-produced proteins are of significant therapeutic value in the pharmaceutical and biochemical industries, particularly when they can be secreted from the host in fully-folded active form. Accordingly, there are further provided the heterologous proteins produced by the Tat secretion pathway using the foregoing methods, and the computer algorithm used to identify the Tat signal sequence and putative substrates.

The present invention relates to a method for secreting and producing a target protein into cell culture broth. More particularly, the invention relates to a microorganism co-transformed with a recombinant expression vector containing E. coli outer membrane protein F (OmpF) and a recombinant expression vector containing a target protein to be secreted into cell culture broth, as well as a method of secreting and producing the target protein into cell culture broth by culturing the microorganism. According to the invention, the target protein can be secreted into cell culture broth in a pure form without fusion with other proteins so that the efficient isolation and purification of the target protein is possible.

The invention discloses a pelB signal peptide mutant capable of improving the protein secretion efficiency and an application of the pelB signal peptide mutant and belongs to the technical field of gene engineering. A sequence of a pelB mutant secretive expression signal peptide is as shown in SEQ ID NO.1. The signal peptide is capable of improving the extracellular enzyme activity of objective protein cyclodextrin glycosyltransferase by 1.6 times. The extracellular protein production capability of recombinant escherichia coli transformed by using the signal peptide is strengthened, and industrialized production is facilitated.

The invention provides alginate lyase, host cells capable of secreting alginate lyase and application of the alginate lyase and the host cells. The alginate lyase has any one of amino acid sequences shown as (I) and (II): (I) an amino acid sequence shown as SEQ ID NO. 1; (II) an amino acid sequence obtained by carrying out substitution, deletion or addition of 1 to 20 amino acid residues on the amino acid sequence shown as SEQ ID NO. 1; the obtained amino acid sequence has the activity of the alginate lyase. According to the alginate lyase, whole genes of the alginate lyase are truncated to reduce the molecular weight, so that the expression amount of the host cells on the alginate lyase is remarkably improved; the secreted alginate lyase has high activity, good stability and strong degradation capability on sodium alginate.

The invention provides an ependymin-like protein derived from a mammal, or its partial peptide or its precursor protein, or a salt thereof; a signal peptide; a DNA coding for the protein, etc. a recombinant vector; a transformant; a method of producing the protein; a pharmaceutical composition comprising the protein, etc. or DNA, an antibody against the protein, etc.; and a method for screening and a screening kit for compounds promoting the function of the protein. The protein, its partial peptide or a salt thereof has physiological activities such as a nerve-extending or nerve-regenerating activity, a gliacyte stimulating activity, and so on. The protein, etc. or the DNA is useful as a therapeutic or prophylactic agent for Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS), dementia or cerebellar degeneration. The antibody against the protein, etc. can be used in the assay of the protein, etc. in a test sample. Furthermore, the protein, etc. is useful as a screening reagent for compounds or their salts capable of promoting the function of the protein.

The present invention discloses an HLA Specific human cytomegalovirus Multi-epitope adenovirus DNA Vaccine of Chinese population, which consists of human cytomegalovirus tandem sequence CTL*Th and adenovirus expression vector pAd5F35, wherein the human cytomegalovirus tandem sequence CTL*Th comprises a nucleotide sequence represented by SEQ ID No.1. The nucleotide sequence comprises the nucleotide sequences of 83 antigen epitopes of 15 encoding proteins of human cytomegalovirus. Furthermore, the sequence covers 14 HLA I allelomorphic gene sites and 7 HLA II allelomorphic gene sites. The coverage percent of the HCMV tandem epitope DNA vaccine representing excellent HLA is 92.07 percent in average, and is used for preventing the forming of HCMV disease in the steps of adhering, duplicating and re-activating of the virus. The in vetro experiment shows that the vaccine of the invention has excellent immune effect and overcomes the defects of inferior immunogenicity and limited immune range of the prior HCMV vaccine.

The invention discloses antibacterial maca spray. The antibacterial maca spray comprises, by weight, 0.5-1.0% of maca extract liquid, 1.0-1.5% of cinnamon extracts, 0.1-0.15% of clove extract liquid, 0.1-0.15% of capsicum annuum extract liquid, 0.1-0.5% of cistanche deserticola extracts, 0.3-0.5% of chlorhexidine acetate and the balance water. The antibacterial maca spray has the advantages that inhibition effects can be realized for hemolytic streptococcus, streptococcus pneumoniae, Escherichia coli bacteria, proteus vulgaris, salmonella paratyphi A, shigella dysenteriae, Escherichia coli, staphylococcus aureus, methicillin-resistant staphylococcus aureus, candida albicans, pseudomonas aeruginosa and the like, the antibacterial maca spray can be used for inhibiting the Escherichia coli, the staphylococcus aureus, the candida albicans and the like and has little skin irritation, and good wetting effects can be realized.

The invention relates to the field of genetic engineering and the field of protein secretion expression and provides a method for constructing a pichia pastoris engineered strain used for preparing dextranase. The method comprises the following steps: (1) designing an oligonucleotides primer and taking lipomyces 1390 genome DNA as a template; cloning to obtain dextranase genes; (2) utilizing a secretion expression carrier pPIC9K to insert the dextranase genes into multi-cloning sites (EcoRI and NotI) of the pPIC9K so as to construct an integral secretion-type expression carrier pPIC9K-139; and(3) linearizing the reconstructed expression carrier pPIC9K-139 so as to realize the high-efficiency secretion expression of the dextranase, and obtaining the reconstructed pichia pastoris engineeredstrain GS115-139 by means of selection. The method successfully realizing the effective expression of the dextranase genes in pichia pastoris, constructs the engineered strain which can efficiently secrete and express the dextranase, and realizes the expansion of a fermentation process of the dextranase.

Construction and expression of synthetic pathways to produce (S) or (R)-3-hydroxybutyrate (3HB) as enantiomerically-pure products by genetically engineering cyanobacterium Synechocystis sp. PCC 6803. Under optimized growth conditions, the pathway employing phaA and phaB from R. eutropha was the most effective, producing up to 533.4±5.5 mg / l (R)-3HB after 21 days photosynthetic cultivation. For the first time, the feasibility and high efficiency of producing 3HB using solar energy and CO2 as sole energy and carbon sources by engineered cyanobacteria is demonstrated.

The present invention is directed to a recombinant E. coli Nissle 1917 (EcN) cell transformed with a nucleic acid coding for a defensin protein or a derivative thereof. The invention is further directed to a pharmaceutical composition comprising this cell and a pharmaceutically acceptable carrier as well as a method of producing a recombinant E. coli Nissle 1917 cell and its use in the treatment of Crohn's disease.

The invention discloses a lignin biological resource utilization method. According to the method, lignin can serve as a fermentation auxiliary substrate, continuous high-density culture is implemented, and the lignin can be degraded into substances such as acetic acid, vanillin, guaiacol, veratryl alcohol and phenolic compounds by fungal complex floras. The lignin can be applied to the fungal complex floras and mass efficient production of active substances of the fungal complex floras. Culture conditions and culture medium components of the fungal complex floras are optimized, the biodegradation rate and the conversion efficiency of the lignin are improved, so that the degradation rate of the lignin is not lower than 40%, and the concentration of a degradation product is 10-100mg / L. An efficient resource utilization method is provided for degradation and conversion of the lignin.

The invention relates to a lactococcus lactis food-level double-screening marker secretory expression vector and a construction method thereof. The vector comprises a melA gene, a nisI fused gene double selection marker containing a P32 promoter, a replicor of a plasmid pWV01, a multiple cloning site, a P32 promoter for secretory expression and a part of sequence of enterococcus faecalis Usp45 secretory protein. The invention also discloses an application of NN1-1 as an exogenous protein expression vector, including recombinant lactococcus lactis of endoglucanase (end-1) constructed for secretory expression of fibrobacter succinogenes with NN1-1 as a vector.

The invention provides construction of TRAIL (Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand) recombinant bacille calmette guerin (rBCG), and relates to a shuttle expression vector comprising a signal peptide fragment of a major secretory antigen Ag85B of BCG and a gene fragment of a TRAIL and a construction method thereof. The obtained shuttle expression vector pMV261-Ag85B-TRAIL is used for constructing rBCGTRAIL, and can be applied to preparation of TRAIL rBCG for treating superficial bladder tumors, preventing postoperative recurrence thereof and preventing tuberculosis. The rBCG has dual functions of TRAIL and BCG, so that cooperative and synergistic actions of the TRAIL and BCG can be better brought into play; rBCG-TRAIL can secrete TRAIL, and the using amount of the rBCG-TRAIL can be lower than that of the BCG under the condition that the same or better immune effect is achieved, so that the toxic or side effect is reduced; and the rBCG-TRAIL can directly secrete TRAIL efficiently on a certain part, so that tumor cells can be killed in cooperation with the rBCG-TRAIL, and high cost caused by the use of a foreign cell factor is avoided.