1024results about How to "Efficient expression" patented technology

The present invention relates to oligopeptide-free cell culture media comprising at least 0.5 mg / L of a polyamine and to methods for cultivating cells in said oligopeptide-free cell culture media comprising at least 0.5 mg / L of a polyamine. The invention also relates to methods for expressing at least one protein in a medium comprising at least 0.5 mg / L of a polyamine and to methods for producing at least one virus in a medium comprising at least 0.5 mg / L of a polyamine.

Provided is a lentiviral vector capable of delivering a nucleotide of interest (NOI) to a desired target site and wherein the NOI encodes the Factor VIII and the Factor VIII is expressed following delivery of the NOI to the desired target site.

The present invention relates to animal protein-free cell culture media comprising polyamines and a plant- and / or yeast-derived hydrolysate. The invention also relates to animal protein-free culturing processes, wherein cells can be cultivated, propagated and passaged without adding supplementary animal proteins in the culture medium. These processes are useful in cultivating cells, such as recombinant cells or cells infected with a virus, and for producing biological products by cell culture processes.

The invention discloses a design method for a distributed hydrological model by using a grid as an analog unit, which is called as ESSI for short. The design method comprises the following steps of: obtainment of a distributed parameter: converting vector data into grid data; generalization of a watershed hydrological process: establishing a universal runoff generating type for the grid; design of a runoff generating process: respectively computing the water-quantity distributing condition of each part according to different prior-period soil water conditions; design of a flow collecting process: respectively designing a Muskingum-Cunge method, a delay algorithm and a riverway segmentation Muskingum method for the flow collecting process computation of the model under different conditions; and model development and integration. The invention not only can finish the watershed hydrological process simulation at an arid region and a humid region, but also can realize the short-term flood forecast and the long-term rainfall-runoff process simulation and prediction of a watershed by using modularization and integration ideas as means, thereby providing scientific reference bases for deeply learning about the physical mechanism of water circulation by people, reducing the drought and water-logging disasters and reasonably developing and utilizing water resources.

A yeast alpha-factor expression system is provided comprised of a truncated leader sequence, containing the alpha-factor signal peptide and one glycosylation site, linked by a processing site to a non-yeast protein-encoding sequence.

Adaptive cooperative web caches construct and maintain routing and forwarding tables for efficient location of replicated web content data, thus reducing the time taken to fetch the requested web content data. The routing and forwarding tables in particular store decomposed web content identifiers and use compression techniques for efficient translation from a user request to the internet protocol address of the cache server where the replicated web content data can be retrieved.

A biodegradable, mixed polymeric micelle used to deliver a selected nucleic acid into a targeted host cell contains an amphiphilic polyester-polycation copolymer and an amphiphilic polyester-sugar copolymer. The polyester-polycation copolymer forms an electrostatic interaction with polyanionic nucleic acids, and the polyester-sugar copolymer directs the micelle-nucleic acid complex to cells in vivo. Additional copolymers with similar properties may also be included. The composition improves delivery efficiency by providing a particulate gene carrier for which particle size and charge density are easily controlled by multivariate means. Various kinds of ligands and other functional compounds may be also be introduced using the composition. The composition may be used in a method for transforming a targeted host cell with a selected nucleic acid.

A context-aware role-based access control system and a control method thereof. The context-aware role-based access control system includes: a context-aware user assignment manager (CAUAM) for performing a role assignment function, a role delegation function, or a role revocation function for a user according to a context of the user, based on a preset context request condition; a context-aware permission assignment manager (CAPAM) for performing a permission modification, a permission restoration, and a personalized permission modification for a permission, which the role has, according to changes in the context of the user; an information repository for storing a user profile and context information; and an access control manager (ACM) for controlling the context-aware user assignment manager, the context-aware permission assignment manager, and the information repository, and processing an access control request. Accordingly, more efficient access control can be achieved in ubiquitous environments where the context of the user dynamically changes.

Compositions for targeted mutagenesis of cell surface receptors for HIV and methods of their use are provided herein. The compositions include triplex-forming molecules that displace the polypyrimidine strand of target duplex and form a triple-stranded structure and hybrid duplex in a sequence specific manner with the polypurine strand of the target duplex. The triplex-forming molecules include a mixed-sequence “tail” which increases the stringency of binding to the target duplex, improves the frequency of modification at the target site, and reduces the requirement for a polypurine:polypyrimidine stretch. Methods for using the triplex-forming molecules in combination with one or more donor oligonucleotides for targeted modification of sites within or adjacent to genes that encodes cell surface receptors for human immunodeficiency virus (HIV) are also disclosed. Methods for ex vivo and in vivo prophylaxis and therapy of HIV infection using the disclosed compositions are also provided.

Disclosed are useful constructs and methods for the expression of proteins using primary translation products that are processed within a recombinant host cell. Constructs comprising a single open reading frame (sORF) are described for protein expression including expression of multiple polypeptides. A primary translation product (a pro-protein or a polyprotein) contains polypeptides such as inteins or hedgehog family auto-processing domains, or variants thereof, inserted in frame between multiple protein subunits of interest. The primary product can also contain cleavage sequences such as other proteolytic cleavage or protease recognition sites, or signal peptides which contain recognition sequences for signal peptidases, separating at least two of the multiple protein subunits. The sequences of the inserted auto-processing polypeptides or cleavage sites can be manipulated to enhance the efficiency of expression of the separate multiple protein subunits. Also disclosed are independent aspects of conducting efficient expression, secretion, and / or multimeric assembly of proteins such as immunoglobulins. Where the polyprotein contains immunoglobulin heavy and light chain segments or fragments capable of antigen recognition, in an embodiment a selectable stoichiometric ratio is at least two copies of a light chain segment per heavy chain segment, with the result that the production of properly folded and assembled functional antibody is made. Modified signal peptides, including such from immunoglobulin light chains, are described.

Some aspect of this invention provide nucleic acid constructs for transgene expression. Some aspects of this invention provide multicistronic nucleic acid constructs, for example, comprising an expression cassette encoding a hairpin RNA and a reporter expression cassette. Some aspects of this invention provide nucleic acid constructs comprising two or more self-complementary nucleic acid sequences, for example, hairpin RNA encoding nucleic acid sequences and AAV inverse terminal repeats. Methods for the use of the constructs in therapy and research are also provided.

The invention belongs to the field of molecular biology and relates to a high-activity T-cell promoter and an application thereof. Particularly, the high-activity T-cell promoter comprises an element 1 and an element 2, wherein the element 1 is an EF1 alpha promoter and / or an EF1 alpha promoter containing an intron; and the element 2 is any one or more of an mCMV promoter, an hCMV promoter and a CD3e promoter. The high-activity T-cell promoter provided by the invention can be used for efficiently expressing a foreign gene in a T cell, has a stable sequence and is free from sequence loss during the transfer process of a prokaryotic cell and a eukaryotic cell. The high-activity T-cell promoter is applicable to driving the high-efficiency expression of the foreign gene, especially a full-length antibody gene, in the T cell during the process of adoptive cellular immunotherapy.

Production of recombinant human serum albumin with rice albuminous cell as biological reactor is carried out by taking rice albuminous cell protein as storage site for recombinant protein, taking rice albuminous specific expression starter and signal peptide, entering inductive recombinant human serum albumin into inner-film system of rice serum albumin cell and storing it in rice albuminous protein. It is cheap, has more yield and higher expression.

The present invention provides a bacterium belonging to the genus Bifidobacterium, by which DNA coding for a protein having an antitumor activity or DNA coding for a protein having the activity of converting a precursor of an antitumor substance into the antitumor substance is delivered to tumor tissues specifically under anaerobic conditions thereby expressing the protein encoded by the DNA, as well as a pharmaceutical composition comprising said anaerobic bacterium.

Compositions for targeted mutagenesis of cell surface receptors for HIV and methods of their use are provided herein. The compositions include triplex-forming molecules that bind to duplex DNA in a sequence specific manner at target sites to form triple-stranded structures. The triplex-forming molecules can be triplex-forming oligonucleotides (TFOs) or peptide nucleic acids (PNAs). The triplex-forming molecules are useful to induce site-specific homologous recombination in mammalian cells when used in combination with donor oligonucleotides. The triplex-forming molecules target sites within or adjacent to genes that encodes cell surface receptors for human immunodeficiency virus (HIV). This binding stimulates homologous recombination of a donor oligonucleotide to cause mutations in HIV cell surface receptor genes that result in one or more deficiencies in the ability of the encoded receptor to bind to HIV and allow its transport into the cell. Methods for ex vivo and in vivo prophylaxis and therapy of HIV infection using the disclosed compositions are also provided.

This invention provides fusion proteins comprising consecutive amino acids which beginning at the amino terminus of the protein correspond to consecutive amino acids present in (i) a cytomegalovirus human MHC-restricted peptide, (ii) a first peptide linker, (iii) a human β-2 microglobulin, (iv) a second peptide linker, (v) a HLA-A2 chain of a human MHC class I molecule, (vi) a third peptide linker, (vii) a variable region from a heavy chain of a scFv fragment of an antibody, and (viii) a variable region from a light chain of such scFv fragment, wherein the consecutive amino acids which correspond to (vii) and (viii) are bound together directly by a peptide bond or by consecutive amino acids which correspond to a fourth peptide linker, wherein the antibody from which the scFv fragment is derived specifically binds to mesothelin. This invention provides nucleic acid constructs encoding same, processes for producing same, compositions, and uses thereof.

The present invention provides a high affinity, antigen-specific, soluble heavy chain-only antibody which: lacks hallmark camelid-related amino acid substitutions and has FR2 substitutions which are not found in antibodies which comprise heavy and light chain; shows increased net hydrophobicity within CDR1 and an increased number of charged amino acids present in CDR3; and comprises one or more amino acid substitutions within the framework β-pleated sheet leading to increased net hydrophobicity within FR1 and increased number of charged amino acids present in FR3. Also provided are VH domains having the same properties, gene segments for their production, methods for their production, transgenic animals and uses of the antibody of the VH domains in therapy.

This invention provides the data network user with an interactive search engine environment that helps him focus on the object of his search. Through a powerful graphical user interface (GUI), the user is provided with terms closely related to what he is looking for, even when he has no prior knowledge of the subject or its terminology. The user is also provided with suggested terms that should be excluded from the search because of the high probability of interfering with the search. The contribution of each term to the search progress is explained to the user. The invention then enables the user to build complex queries combining the related terms in a visual and simple way that facilitates the search process and saves time. This invention allows the user to create a detailed, precise, and longer search query then is customarily executed today

The present invention relates to animal protein-free cell culture media comprising polyamines and a plant and / or yeast-derived hydrolysate. The invention also relates to animal protein-free culturing processes, wherein cells can be cultivated, propagated and passaged without adding supplementary animal proteins in the culture medium. These processes are useful in cultivating cells, such as recombinant cells or cells infected with a virus, and for producing biological products by cell culture processes.

Improved molecular vaccines comprise nucleic acid vectors that encode a fusion polypeptide that includes polypeptide or peptide physically linked to an antigen. The linked polypeptide is one that (a) promotes processing of the expressed fusion polypeptide via the MHC class I pathway and / or (b) promotes development or activity of antigen presenting cells, primarily dendritic cells. These vaccines employ one of several types of nucleic acid vectors, each with its own relative advantages: naked DNA plasmids, self-replicating RNA replicons and suicidal DNA-based on viral RNA replicons. Administration of such a vaccine results in enhance immune responses, primarily those mediated by CD8+ cytotoxic T lymphocytes, directed against the immunizing antigen part of the fusion polypeptide. Such vaccines are useful against tumor antigens, viral antigens and antigens of other pathogenic microorganisms and can be used in the prevention or treatment of diseases that include cancer and infections.

A context-aware role-based access control system and a control method thereof. The context-aware role-based access control system includes: a context-aware user assignment manager (CAUAM) for performing a role assignment function, a role delegation function, or a role revocation function for a user according to a context of the user, based on a preset context request condition; a context-aware permission assignment manager (CAPAM) for performing a permission modification, a permission restoration, and a personalized permission modification for a permission, which the role has, according to changes in the context of the user; an information repository for storing a user profile and context information; and an access control manager (ACM) for controlling the context-aware user assignment manager, the context-aware permission assignment manager, and the information repository, and processing an access control request. Accordingly, more efficient access control can be achieved in ubiquitous environments where the context of the user dynamically changes.

The invention mainly aims to provide a porcine circovirus type 2 (PCV2) subunit vaccine and a preparation method thereof. Particularly, a baculovirus vector expression system is utilized to express a large amount of recombinant open reading frame type 2 (ORF2) protein in insect cells, so that the subunit vaccine with good immunity effect is developed. A bac-to-bac baculovirus expression system is adopted to perform whole gene amplification on the porcine circovirus type 2 ORF2 gene, and a melittin signal peptide nucleotide sequence is introduced into a terminal 5', so that the recombinant baculovirus of an open reading frame containing the melittin signal peptide nucleotide sequence and a PVC2ORF2 gene sequence is established, wherein the infected insect cell expresses the recombinant ORF2 protein with efficient and high PCV2 antigenicity; and thus the subunit vaccine containing the PCV2 recombinant ORF2 protein is prepared. The inoculation experiments of piglets show that the subunit vaccine has a good immunity protection effect.

The invention provides a DNA (deoxyribonucleic acid) sequence used for circular RNA (ribonucleic acid) expression and a vector containing the DNA sequence used for circular RNA expression. The vector is inserted in a eukaryotic expression vector, a lentiviral vector, an adenovirus vector or a retroviral vector to form a series of special expression vectors for circRNA. The DNA sequence and the corresponding expression vector have the best effects, are simple and convenient to operate and are stable in results and efficient in expression (the circRNA expression level is increased by more than a hundredfold). The sequence and the corresponding vector can be widely applied to expression of various circRNA, provide a powerful research tool for the functions and mechanisms of circRNA and provide theoretical support for research and development of further determination of circRNA molecules as novel markers and disease treatment targets.

The invention discloses a Bayesian network-based regional heat supply model predictive control system and method. The method includes the following steps that: step S1, physical layer heat network data sensing is performed: historical data are acquired from a source side, a network side and a building side in real time and are updated; step S2, a Bayesian network is constructed according to the historical data and on the basis of prior knowledge, and the load demand of a thermal station and the building side are predicted through the Bayesian network; step S3, the real-time control parametersof a secondary side, a primary side and the source side are obtained through the inference of the Bayesian network according to the load demand of the building side and on the basis of the historicaloperation data and real-time data; and step S4, time characteristic curves of source side adjustment, network side adjustment and building side adjustment are established according to the historical operation data and pipe network topology structures, and source side adjustment, network side valve and building side electric valve adjustment strategies are determined, control operations are performed according to the strategies, so that the hysteresis of heat network adjustment can be eliminated, real-time supply and demand balance can be met, and the accurate on-demand heat supply of a heat user side can be realized.

The present invention relates to compositions and methods for creation of vector nucleic acid sequences (e.g., retroviral nucleic acid sequences) that comprise two or more exogenous nucleic acid sequences that encode highly homologous (e.g., identical) polypeptide sequences, yet wherein at least one of the exogenous nucleic acid sequences has been mutated using degenerate codons for purpose of reducing homology between the two or more exogenous nucleic acid sequences while maintaining the encoded polypeptide sequence. Preferred nucleic acid sequences include those encoding multi-chimeric immune receptor (CIR) genes. Specific nucleic acid sequences of such CIR genes are also disclosed.

The present invention provides an oncolytic virus comprising nucleotide sequence(s) encoding one or more immune checkpoint modulator(s). It also concerns a pharmaceutical composition comprising effective amount of said oncolytic virus and, eventually, a pharmaceutically acceptable vehicle and its use for treating proliferative diseases such as cancers.