571results about How to "Improve expression efficiency" patented technology

A significantly increased amount of a monoclonal antibody is obtained from the culture medium of recombinant hybridoma prepared by introducing genes encoding a protein identical to the immunoglobulin heavy chain polypeptide of the specific monoclonal antibody into an immortalized B cell (hybridoma) producing the monoclonal antibody.

The invention discloses a CAR-T transgene vector based on replication defective recombinant lentivirus. The CAR-T transgene vector comprises an original nuclear replicon pUCOri sequence, a resistance gene AmpR sequence containing ampicillin, a virus replicon SV40 Ori sequence, a lentivirus packaging cis element, ZsGreen1 green fluorescent protein, an IRES ribosome binding sequence, a human EF1 alpha promoter , a chimeric antigen receptor of second-generation CAR or third-generation CAR and a regulating element, wherein the original nuclear replicon pUCOri sequence is used for plasmid replication; the resistance gene AmpR sequence is used for massively proliferating target strains; the virus replicon SV40 Ori sequence is used for enhancing replication in eukaryocyte; the lentivirus packaging cis element is used for lentivirus packaging; the ZsGreen1 green fluorescent protein is used for expressing green fluorescent for eukaryocyte; the IRES ribosome binding sequence is used for jointly transcribing and expressing protein; the human EF1 alpha promoter is used for conducting eukaryotic transcription on antigen receptor genes; the chimeric antigen receptor is used for forming the second-generation CAR or the third-generation CAR integrating recognition, transfer and start; the regulating element is used for enhancing expression efficiency of transgenes and used after eWPRE-enhanced type woodchuck hepatitis b virus is transcribed. Besides, the invention further discloses a construction method and application of the vector. By means of the CAR-T transgene vector and the construction method and application of the vector, secretion of cell factors and an in vitro killing effect of CAR-T cells can be remarkably improved, and the clinical treatment effect is remarkable.

The invention provides human extracellular adhesive proteins (EXADH) and polynucleotides which identify and encode EXADH. The invention also provides expression vectors, host cells, antibodies, agonists, and antagonists. The invention also provides methods for diagnosing, treating or preventing disorders associated with expression of EXADH.

The invention discloses a facial feature recognition method and system based on the multi-region characteristic and metric learning. The method comprises the steps that convolution neural network parameters of the corresponding location and scale are obtained through the multi-scale facial area training, and corresponding facial area features are extracted according to the neural network parameters; the above features are filtered to obtain the high dimensional facial features; metric learning is conducted according to the high dimensional facial features, the after defined loss function of the feature expression is obtained through the dimension reduction processing of the features, a network model of the metric learning is obtained through the training of the loss function; the images to be recognized are inputted into the network model, the facial features are dimension reduced using the Euclidean distance in order to be recognized. In the method, multiscale is used to select multiple areas, the convolutional neural networks are trained, and the expression skills of the characteristics are improved. Meanwhile, through the selection of the acquired multi-scale features, the efficiency of expression of characteristics is improved, and the accuracy of face recognition is effectively improved.

The invention provides human extracellular signaling molecules (EXCS) and polynucleotides which identify and encode EXCS. The invention also provides expression vectors, host cells, antibodies, agonists, and antagonists. The invention also provides methods for diagnosing, treating, or preventing disorders associated with expression of EXCS.

The invention provides a construction method of a human papilloma virus resistant HPV polyvalent vaccine and application thereof.

The invention discloses an anti-BCMA chimeric antigen receptor, an encoding gene, a recombinant expression vector and an establishing method and application of the anti-BCMA chimeric antigen receptor, the encoding gene and the recombinant expression vector. The receptor comprises a CD8 leader chimeric receptor signal peptide, a BCMA single-chain antibody heavy chain VH, an Optimal Linker C, a BCMA single-chain antibody light chain VL, a CD8 Hinge chimeric receptor hinge, a CD8 Transmembrane chimeric receptor transmembrane domain, a CD137 chimeric receptor co-stimulatory factor and a TCR chimeric receptor T cell activating domain which are sequentially connected in series. In addition, the invention further discloses the encoding gene and the recombinant expression vector of the anti-BCMA chimeric antigen receptor and the establishing method and application of the encoding gene and the recombinant expression vector. The secretion of cell factors and the cytotoxicity in vitro of CAR-T cells can be remarkably improved, and the clinical treatment effect is outstanding.

The invention relates to drug-loading nano-micelles, and a preparation method and an application thereof. According to the to drug-loading nano-micelles, a polyethylene glycol derivative-polylysine-polyleucine amphiphilic triblock copolymer is adopted as a carrier, and self-assembly is carried out in a water solution, such that nano-micelles with a three-layer structure transferring gene and medicine are formed. With the hydrophobic effect between polyleucine, the copolymer forms a core of the micelles in water, such that a hydrophobic medicine docetaxel is entrapped. With polylysine, the nano-micelles are positively charged, such that negatively charged anti-apoptotic protein small interfering RNA can be bonded. Polyethylene glycol is used for protecting the stability of the nano-micelles loading the medicines and gene, and a cycle time of the nano-micelles in vivo is prolonged. The nano-micelles are used for loading both an anti-apoptotic protein small interfering RNA gene medicine and a docetaxel medicine, such that the gene treatment medicine and the chemotherapeutic medicine cooperates in treating cancer, and a synergetic effect of the two medicines can be developed. Therefore, a direction is provided for better treatment of breast cancer.

The invention provides human drug metabolizing enzymes (DME) and polynucleotides which identify and encode DME. The invention also provides expression vectors, host cells, antibodies, agonists, and antagonists. The invention also provides methods for diagnosing, treating, or preventing disorders associated with aberrant expression of DME.

The present invention provides an expression vector for the production of proteins and peptides comprising a promoter operably linked to gene of interest, TPL and VA genes I and II, matrix attachment regions (MARs) / SARs, and antibiotic marker. The vector is transfected in suitable host cell.

The invention relates to a recombinant nucleic acid molecule for transcribing circular RNA and application of the recombinant nucleic acid molecule in protein expression. Specifically, the invention relates to the recombinant nucleic acid molecule for transcribing circular RNA, a recombinant expression vector, linear RNA, circular RNA, a recombinant host cell, a pharmaceutical composition and a method for preparing protein. The recombinant nucleic acid molecule is transcribed to form circular RNA containing a specific IRES element, the IRES element can improve the protein expression level of the circular RNA in eukaryotic cells, efficient and durable expression of protein is achieved, and the recombinant nucleic acid molecule has important application values for preparing mRNA infectious disease vaccines, therapeutic mRNA tumor vaccines and mRNA-based dendritic cell tumor vaccines, and in the fields of gene therapy based on mRNA, chimeric antigen receptor T cell therapy based on mRNA,protein supplement therapy and the like.

The invention relates to a recombined fusion protein, in particular discloses a recombined long-acting glucagons peptide analogue and a preparation method thereof. The long-acting glucagons peptide analogue is polypeptide by directly connecting duodenin analogue exendin-4 and human serum albumin, and no connecting peptide exists between the exendin-4 and the human serum albumin. The long-acting glucagons peptide analogue greatly prolongs the half-life of the exendin-4 and is clinically used to reduce the frequency of injection administration, lighten the suffering of patients, improve the curative effect and decrease the hospitalization cost.

The invention relates to a PCV2 (porcine circovirus 2) ELISA (enzyme-linked immuno sorbent assay) antigen detection kit as well as a preparation method and applications thereof, wherein the detection kit comprises an elisa plate of a polyclonal antibody of peridium anti-PCV2-Cap (nucleocapsid) protein, seal liquids, sample diluent, an antigen standard product, a second antibody of a monoclonal antibody of HRP marked anti-PCV2-Cap protein, a concentrated washing liquid, an enzyme substrate solution A, an enzyme substrate solution B and a stop solution, wherein the antigen standard product is purified reconstructed PCV2-Cap protein. The specificity of the kit provided by the invention achieves 100%, and the sensitivity is as high as 4ng / ml, and the kit can be used for swinery PCV2 antigen detection and PCV2 vaccine product quantitative detection.

A vector which is characterized in containing each of the following elements:(1) a promoter which shows its action in the host to be used;(2) regulatory region for regulating the action of the promoter of (1); and(3) a region which codes for the 5'-untranslated region derived from cold-shock protein gene mRNA or a region which codes for the region where substitution, deletion, insertion or addition of at least one base is applied to the said untranslated region.

The mutations D188V, V1353L, I1656M in the neuronal gene sodium-channel alpha1-subunit, SCN1A, are disclosed. The methods of using their associated polypeptides for treating sodium channel dysfunction disorders including generalized epilepsy are also disclosed.

The invention belongs to the technical field of biological medicine, relating to an HIV-1gp120 gene consensus sequence optimized by codons and a gp120 nucleic acid vaccine. In the invention, a Chinese HIV-1 epidemic strain (A / E recombinant subtype, B / C recombinant subtype and ThB subtype) envelope protein gp120 consensus amino acid sequence is obtained by applying multi-sequence comparative analysis; a gp120 gene segment encoding the consensus amino acid sequence is prepared by using an artificially synthesized method, and the gene is optimized by the codons and combines preferences of mammalian cells and escherichia coli codons; and on the basis, HIV-1gp120 nucleic acid vaccine consisting of the gp120 gene and eukaryotic expression vector pJW4303 is constructed. The nucleic acid vaccine can be applied to animal and human immunization and expression and production of the gp120 protein.

The invention relates to a 2 type subunit vaccine for a porcine circovirus as well as a preparation method and application thereof. A recombinant bacilliform virus contains double promoters (a polyhedrin promoter and a P10 promoter), a coding gene of a Cap protein with double copying can be expressed, and the expression efficiency of the protein is obviously enhanced; moreover, the Cap protein expressed by an inserted foreign gene does not contain an excess sequence, virus-like particles (VLPs) can be effectively formed, and the immunogenicity of an expressed protein is enhanced; furthermore, a produced antigen has high content; and according to the 2 type subunit vaccine for the porcine circovirus, which is disclosed by the invention, the productivity ratio and the quality of a viral protein of the 2 type subunit vaccine for the porcine circovirus are obviously enhanced, and a prepared vaccine composition has the advantages of stable and persistent immune effect, high safety and the like.

The present invention relates to DNA constructs that can produce antimicrobial materials efficiently from microorganisms and the preparation method thereof. The present invention also relates to the useful vector for the DNA construct. The DNA construct according to the present invention comprises a first gene coding for entire, a part of or a derivative of purF gene and a second gene coding for antimicrobial peptide. According to the present invention, antimicrobial peptides can be mass-produced by the following steps: preparing an expression vector containing a DNA construct comprising a first gene coding for an entire, a part of or a derivative of purF gene and a second gene coding for antimicrobial peptide; transforming the bacterial host cells with the above-mentioned vector, culturing the transformed cell to express the above-mentioned DNA construct; and recovering the above antimicrobial peptide.

The invention discloses a method for producing a porcine parvovirus antigen and its product. The method comprises the following steps: porcine parvovirus capsid protein VP2 gene or optimized VP2 gene is cloned in a baculovirus carrier so as to obtain a transfer expression carrier; the constructed transfer expression carrier and baculovirus DNA are carried out cotransfection to obtain recombined baculovirus; the recombined baculovirus is used to infect insect host and cell; the infected insect host is cultured to express corresponding porcine parvovirus capsid protein; and the expressed antigen is ingathered and purified so as to obtain the porcine parvovirus antigen. The method adopts a baculovirus expression system to make safe and efficient porcine parvovirus antigen capsid particles in a domestic silkworm bioreactor; the prepared purified antigen by the method has high safety, and can be directly produced to vaccines for animal immunity. The method for producing porcine parvovirus antigen has the advantages of high expression efficiency, high immunization activity of the expressed antigen, low production cost, large scale production realization and the like.

The invention relates to a fusion protein MBP-NAP. The amino acid sequence of the fusion protein is shown as SEEQ ID NO.1. Moreover, the invention also discloses a preparation method of the fusion protein MBP-NAP. The method has the characteristics that: the method is easy and convenient to operate, has a mild condition and the like. Th1 cellular immunity of the fusion protein MBP-NAP of the invention is enhanced, so that the secretory volumes of related cell factors IL-2 and IFN gamma are increased remarkably and Th1 immunity is enhanced.

The invention discloses recombinant pichia pastoris for heterogenous high level secretory expression of a rhizomucor miehei lipase. The recombinant pichia pastoris is obtained by introducing a self leader peptide gene for encoding the rhizomucor miehei lipase by employing a molecular biology technology, then optimizing a copy number of an objective gene, and constructing a vector which can be expressed in pasteur pichia pastoris. Researches show that: the introduction of the self leader peptide greatly increases the secretion amount of the objective lipase, a pichia pastoris recombinant of a lipase gene (pro-rml) containing two copies has highest enzyme activity, and the lipase Pro-RML has higher activity of hydrolysis of tri-acylglycerol.

The invention discloses a single-frame resolution ratio reconstruction method based on sparse coding and combined mapping. The method comprises the following steps that: processing an initial high-resolution training set image to obtain an expanded high-resolution feature block sample and an interpolated medium and high resolution feature block sample; training an obtained feature sample, obtaining a dictionary atom as a clustering center, and clustering samples by the center; according to a corresponding relationship among different resolutions, solving the mapping matrix of each cluster; on the basis of the low-resolution image processing way of the training set, processing an input low-resolution test image, and solving the sparse coefficient of the low-resolution test image by the dictionary atom obtained by training; taking the sparse coefficient as a weight, taking each mapping matrix obtained by clustering as a combined element, carrying out matched combination to obtain a mapping relationship required by image reconstruction, and directly multiplying the mapping matrix by an interpolated medium and high resolution feature block to obtain a high-resolution feature block; and carrying out overlap removal and block fusion, and adding original low-frequency information to obtain the reconstructed high-resolution image.

A promoter, an intron and a terminator by which a useful foreign gene can be efficiently expressed in algae and which are thus useful in breeding algae using genetic engineering techniques.

The invention discloses an oligonucleotide aptamer for prostate cancer targeting gene, a delivery carrier, a delivery system and a preparation method thereof. The delivery carrier is made of polycationic polymer material, hydrophilic polymer and Based on the oligonucleotide aptamer, the three are covalently linked, wherein the molar ratio of the hydrophilic polymer to the polycation polymer is 1-25:1, and the oligonucleotide aptamer The molar ratio of the body to the polycation polymer material is 1-5:1; the delivery system is a nanoscale gene delivery system formed by delivery carriers and miRNA, siRNA or plasmid DNA. The delivery carrier and the delivery system of the present invention can effectively improve the gene transfection efficiency and expression efficiency on prostate cancer cells expressing prostate specific membrane antigen, compared with the gene delivery system constructed by targeting the head group used in the prior art , which has the advantages of stronger stability and significantly improved targeting effect.

An isolated nucleic acid molecule encoding a mutant alpha subunit of a mammalian voltage-gated sodium channel, wherein a mutation event selected from the group consisting of point mutations, deletions, insertions and rearrangements has occurred and said mutation event disrupts the functioning of an assembled sodium channel comprising this mutated subunit so as to produce an epilepsy phenotype, polypeptides encoded by said nucleic molecule and uses of these molecules in preparing animal models and in diagnostic applications

The invention discloses an optimized SARS coronary virus spike protein gene, which codes the SARS coronary virus S protein or immunogenicity fragment, and has 77+ / -3% homology with SARS coronary virus S protein coding sequence(such as SEQ ID NO: 1). The invention also provides vector, host cell and vaccine composition containing a optimized S protein coding sequence, whose expression efficiency is higher in mammal cell, So can stimulate the immune response in mammal to SARS.

This invention provides a recombinant protein A gene, whose nucleotide acid sequence is shown in SEQ ID NO.1, and the recombinant protein A coded by the gene. This invention also provides recombinant expression carrier PET32a-P for expressing the recombinant protein A gene, E. coli BL21 / DE3 converted by the gene, and the method for preparing recombinant protein A. Recombinant protein A can be used for antibody detection, separation and purification, and can constitute affinity chromatography medium together with chromatography medium carrier. Recombinant protein A has such advantages as high protein combination specificity, high affinity and high purification efficiency, and has obviously increased dynamic adsorption capacity compared with commercialized protein A sepharose 4 fast flow.

The invention discloses a method for producing a body interleukins29 and restructuring the engineering bacteria IL-29, which comprises artificial synthetic primers, PCR augmenting mature peptides of IL-29 coding areas, the construction of IL-29 pronucleus expression carrier pET44-IL-29, transforming into colibacillus to restructure engineering bacteria IL-29, culturing the engineering bacteria to highly effective express IL-29, and purifying IL-29. In the invention, adopting a less dependent interconnection reaction clone process (LIC) to precise and quick-speed get the mature peptides coding areas of IL-29, adopting the terminator codon of colibacillus preference to increase the expression efficiency, one-step purifying using a S protein affinity chromatography column to get the very purified production.