42 results about "Bicistronic mrna" patented technology

Compositions and methods relating to microRNA (miRNA) technology are disclosed. In particular, microRNA (miRNA) expression vectors and methods for the treatment of sensory disorders, e.g., for the treatment of hearing loss, are described.

The invention relates to a bicistronic mRNA coexpression gene transporter and a preparation method thereof. The gene order of the gene transporter is SEQ ID No. 15; the gene transporter from 5' end to 3' end comprises bovine Nuclear matrix binding region MAR, artificially constructed combined promoter CAG, Profilin gene FSTN, internal ribosome entry site (IRES), green fluorescent protein AcGFP gene and Rabbit globin poly A signal region; the preparation method comprises the following steps: constructing a carrier pCAG-IRES2-AcGFP1; obtaining FSTN gene and inserting the pCAG-IRES2-AcGFP1 carrier; obtaining the sequence of MAR and inserting the pCAG-IRES2-AcGFP1 carrier; constructing a plasmid carriers so as to obtain the bicistronic mRNA coexpression gene transporter. The gene transporter is not only pure and safe gene transporter but also realizes dual-gene coexpression in any combination, achieves the purpose of multi-gene coexpression through repeated utilization of IRES, and provides new thinking and route for improving the shape of dual or multiple gene control such as economic character.

The invention discloses a bicistronic mRNA (messenger ribonucleic acid) expression vector which is suitable for cells of mammals and can realize high-efficient expression of exogenous genes and an application thereof. The bicistronic mRNA expression vector comprises connecting elements in the 5'-3' direction, namely (1) an open reading frame 1: containing the sequences of an early promoter / enhancer and an introne of cytomegalovirus, multiple clone sites and the sequence of a bovine growth hormone transcription terminator, which are sequentially arranged; (2) an open reading frame 2: containing the sequences of the early promoter / enhancer and the introne of the cytomegalovirus, the multiple clone sites and the sequences of the bovine growth hormone transcription terminator and a simian virus promoter, which are sequentially arranged; and (3) screening marker genes and the sequence of a simian virus 40 (SV40) terminator. The bicistronic mRNA expression vector has the high-efficient promoter, effective transcription termination signals, screening and gene amplification markers and two open reading frames, and can improve the expression efficiency of the exogenous genes and reduce the production cost.

The invention discloses methods for detecting bicistronic mRNA viruses of a scylla serrata and kits thereof. A poly-anti-mono monoclonal-antibody and double-antibody sandwich ELISA method and a double-antibody and monoclonal-antibody sandwich ELISA method are adopted for detecting the bicistronic mRNA viruses of the scylla serrata by using monoclonal antibody combined with MCDV specificity. Both the poly-anti-mono monoclonal-antibody and double-antibody sandwich ELISA method and the double-antibody and monoclonal-antibody sandwich ELISA method can sensitively and specifically detect the MCDV masculine in the haemolymph of the virus-infected crabs with the detectable rate of 82 to 92 percent, while the double-antibody and monoclonal-antibody sandwich ELISA method has better sensitivity and specificity on the detection of the bicistronic mRNA viruses of the scylla serrata. The two methods of the invention have the advantages that: corresponding kits can be prepared by the two methods, so the MCDV can be detected with fast speed, high efficiency, sensitivity, specificity and stability, and the methods have instructive meaning for detection in production and future scientific researches.

It is intended to provide a method of screening a so-called Tc inducible gene expressing cell line, in which the expression of a gene of interest is regulated depending on the presence / absence of a Tc compound regardless of the type of cell, easily with high efficiency. A gene-of-interest expression vector for transfecting a gene of interest into the genome of a host cell and a TA expression vector that expresses a transactivator that is switched to be bound or not to be bound to a tet operator sequence depending on the presence or absence of tetracycline are prepared. The gene-of-interest expression vector is a vector with a bicistronic regulatory sequence arranged downstream of the tet operator sequence and promoter sequence and between a gene-of-interest coding sequence and a selection marker coding sequence. These vectors are transfected into a host cell, and cell lines in which the selection marker of the gene-of-interest expression vector is expressed are selected. Thus, inducible gene expressing cell lines can be obtained in which the expression of a gene of interest can be controlled with Tc.

The invention discloses a primer group for detecting the reovirus and bicistronic virus of scylla serrata, comprising a reovirus forward primer ReoF, a reovirus reverse primer ReoR, a bicistronic virus forward primer DicF and a bicistronic virus reverse primer DicR, wherein each primer is specifically as follows: ReoF: 5-ACTCATAGAGCAGTCATGGG-3; ReoR: 5-ATATCGTCAGAATGTCGTTC-3; DicF: 5-GGATACTATGGATGATGTTTC-3; and DicR: 5-ACAAAATACCAGATAAAGCAA-3. The invention further discloses a kit containing the primer group and a detection method. The primer group, the kit and the detection method are shortin detection time, strong in specificity, high in detection sensitivity, and capable of simultaneously detecting the reovirus and bicistronic virus of scylla serrata.

An embodiment of the invention provides bicistronic chimeric antigen receptor (CAR) amino acid constructs. Nucleic acids, recombinant expression vectors, host cells, populations of cells, and pharmaceutical compositions relating to the CAR constructs are disclosed. Methods of detecting the presence of cancer in a mammal and methods of treating or preventing cancer in a mammal are also disclosed. Methods of making the CAR constructs are disclosed.

A tissue culture system for production of infectious hepatitis C virus is described. In particular, the invention provides recombinant monocistronic and bicistronic genomic constructs for production of virus, including constructs for production of wild-type HCV type 2a strain JFH1 and constructs for production of chimeric viruses comprising HCV proteins from strain JFH1 and a second HCV isolate. Constructs of the invention also include a reporter gene to facilitate measurement of RNA replication and viral infectivity in cultures. The cell culture system may also include various factors that improve viral replication or infectivity. In addition, a neutralization assay using HCV grown in cell culture is described.

The present invention discloses a series of eukaryotic expression vectors utilizing the reduction of transcription read-through events to create stable and high-yield cell lines for recombinant protein expression. The vectors comprise more than one polyadenylation signal or one or more polyadenylation signals plus other DNA fragment which is known to enhance transcription termination to control the expression level of selection marker, with the configuration to transcribe the minimal level of full-length bicistronic mRNA to express the selection marker, which can be used to create stable cell lines at high expression levels, without the need for drug selection or drug mediated gene amplification.

This invention relates to bicistronic flavivirus vectors, methods of using such vectors in the prevention and treatment of disease, and methods of making such vectors.

A bicistronic construct of p53 and p14ARF, vectors and other delivery vehicles which include the bicistronic construct, and methods of treating cancer using the vectors and bicistronic construct.

The invention discloses a dicistronic specific DNA utilizing a lacZ alpha oligopeptide encoding gene as a second gene encoding frame and application of the dicistronic specific DNA in expressing a target gene. A specific DNA molecule provided by the invention has a bicistronic structure, and sequentially comprises the following components from upstream to downstream: a lac promoter, a lac control region, a first ribosome bind site, a gene encoding frame I, a second ribosome bind site, a gene encoding frame II and a terminator; the last nucleotide of the gene encoding frame I is connected with a first nucleotide of the second ribosome bind site; and the gene encoding frame II encodes lacZ alph oligopeptide formed by the first to nth amino acid residues from the N terminal of lacZ, wherein n is a natural number within a range of 50-150. The specific DNA molecule provided by the invention has dual capabilities of judging whether an extraneous target gene in the gene encoding frame I is expressed or not and judging the expression amount of extraneous target genes by directly observing without special steps, and has great application value.

The present invention relates to the production of structural proteins and virus-like particles (VLPs) of flaviviruses and hepatitis C. Production is through use of a single expression cassette comprising a viral structural gene, a furin encoding gene and a bicistronic expression element such as an internal ribosome entry site (IRES) between the viral and furin genes. Both the viral gene and furin gene can include a partial capsid encoding sequence acting as a signal peptide to co-locate the viral protein and furin and to act as a membrane anchor for the viral protein and furin. A separate expression cassette comprising a non-structural viral gene can be combined with the initial cassette. The structural proteins and VLPs can be used in vaccines and in the treatment of viral infections.

The invention relates to a 2A peptide, a bicistronic expression vector, a recombinant protein expression system and application. The invention provides the 2A peptide (SEQ ID NO: 1 or SEQ ID NO: 3) and provides the bicistronic expression vector containing an encoding gene of the 2A peptide and a construction method for the bicistronic expression vector; and by applying the 2A peptide to expressionof a recombinant protein through constructing the recombinant protein expression system containing the 2A peptide, efficient expression of 2 target genes in cells of mammals can be achieved. Tests prove that on an equal footing, compared with other 2A peptides, the E2A-and-F2A-based recombinant protein expression system provided by the invention can be used for remarkably improving a stable expression level of recombinant proteins of mammalian cells and can be extensively applied to preparation of various target proteins, particularly antibodies. A foundation is settled for improving an expression system for the mammalian cells, further increasing the expression level of the recombinant proteins, improving stability of cell lines and reducing production cost.

The invention relates to a macrobrachium rosenbergii bicistronic messenger ribonucleic acid (mRNA) viral genome complete sequence and an application thereof, which belong to the technical field of virus genomics. The invention provides the macrobrachium rosenbergii bicistronic mRNA viral genome complete sequence shown as SEQIDNO: 1. In addition, the invention provides a macrobrachium rosenbergii bicistronic mRNA viral specific primer, a probe or a reagent kit. The macrobrachium rosenbergii bicistronic mRNA viral genome complete sequence and the application lay the reliable foundation to the purposes of macrobrachium rosenbergii dicistrovirus (MRDV) detection, MRDV infection path study and the like.

Provided is a gene targeting vector capable of highly efficient gene targeting.A gene targeting vector in which a DNA sequence allowing for bicistronic expression is present 5′ upstream of a selection marker. A method for producing a gene targeting vector, comprising linking a DNA fragment homologous to a 5′ upstream region of a target site, a selection marker having a DNA sequence allowing for bicistronic expression present 5′ upstream thereof, and a DNA fragment homologous to a 3′ downstream region of the target site.

The present invention discloses a series of eukaryotic expression vectors utilizing the reduction of transcription read-through events to create stable and high-yield cell lines for recombinant protein expression. The vectors comprise more than one polyadenylation signal or one or more polyadenylation signals plus other DNA fragment which is known to enhance transcription termination to control the expression level of selection marker, with the configuration to transcribe the minimal level of full-length bicistronic mRNA to express the selection marker, which can be used to create stable cell lines at high expression levels, without the need for drug selection or drug mediated gene amplification.

The present invention relates to a bicistronic expression vector for silencing a gene specifically in astrocytes and neurons, comprising two expression cassettes comprising a first and a second silencer sequence, respectively, wherein the expression of said first silencer sequence within astrocytes is regulated by an astrocyte-specific promoter and the expression of said second silencer sequence within neurons is regulated by a neuron-specific promoter. In a preferred embodiment, said first and second silencer sequences are SOD1 silencer sequences. Pharmaceutical composition comprising said bicistronic vector and the use of the same in the treatment of motoneuron diseases are further described.

The present invention relates to a bicistronic expression vector for silencing a gene specifically in astrocytes and neurons, comprising two expression cassettes comprising a first and a second silencer sequence, respectively, wherein the expression of said first silencer sequence within astrocytes is regulated by an astrocyte-specific promoter and the expression of said second silencer sequence within neurons is regulated by a neuron-specific promoter. In a preferred embodiment, said first and second silencer sequences are SOD1 silencer sequences. Pharmaceutical composition comprising said bicistronic vector and the use of the same in the treatment of motoneuron diseases are further described.

The invention discloses a Bcl2 mutant capable of promoting larger gene expression and its application. The Bcl2 mutant of the present invention is that the 190th codon of the mouse Bcl2 gene is mutated from GGA to a stop codon TGA, or the 193rd codon of the human Bcl2 gene is mutated from GGA to a stop codon TGA. In the present invention, by mutating the 568th base (the 577th position of the human Bcl2 gene), G is changed to T, and the mutated phosphorylated Bcl2 and the objective recombinant protein gene are constructed on a bicistronic expression vector. Promote the expression of the target recombinant protein gene larger than 4.3kb.

The invention relates to a macrobrachium rosenbergii bicistronic messenger ribonucleic acid (mRNA) viral genome complete sequence and an application thereof, which belong to the technical field of virus genomics. The invention provides the macrobrachium rosenbergii bicistronic mRNA viral genome complete sequence shown as SEQIDNO: 1. In addition, the invention provides a macrobrachium rosenbergii bicistronic mRNA viral specific primer, a probe or a reagent kit. The macrobrachium rosenbergii bicistronic mRNA viral genome complete sequence and the application lay the reliable foundation to the purposes of macrobrachium rosenbergii dicistrovirus (MRDV) detection, MRDV infection path study and the like.

The invention discloses a bicistronic expression carrier, an expression system, a preparation method and an application, and belongs to the technical field of genetic engineering. The expression vector contains the sequence of the nuclear matrix binding region, the sequence of the inner ribosome entry site and the screening marker. By inserting the target gene between the promoter and the IRES sequence, a bicis sequence of the target gene-IRES sequence-selection marker gene can be formed. anti subsequence. The structure of the expression vector is: MAR-promoter-target gene-IRES sequence-selection marker-PolyA-MAR. The vector uses a promoter to realize the simultaneous expression of the target gene and the screening marker, which can reduce the false positive cell clones caused by the existence of different expression cassettes, improve the screening rate of positive cell clones, and overcome the unbalanced expression of the target gene and the screening marker in traditional vectors Question: The inclusion of MAR sequences on the vector can overcome transgene silencing and achieve efficient and long-term expression of transgenes in host cells.

The present invention relates to the dual, independent cistron expression system in a single vector for the production of protein of interest proteins and peptides expressed as insoluble inclusion bodies formed in the bacteria E. coli. The present invention also provides the process for the expression of protein of interest using said bicistronic vector.

The invention relates to a virus construct for treating liver cancer and an application and construction method of the virus construct. The invention also relates to a novel oncolytic virus for individualized targeted treatment on liver cancer and a construction method thereof. The construct of the present invention comprises an AFP promoter, an RGD-4C gene, an IL-12-p40 gene, an IRES gene, an IL-12-p35 gene, and optionally a glutamyl transpeptidase gene and a tumor-associated antigen gene in an effective ligation manner.

This invention relates to bicistronic flavivirus vectors, methods of using such vectors in the prevention and treatment of disease, and methods of making such vectors.

The invention provides a bicistronic DNA vaccine for grouper viral nervous necrosis, which is a recombinant eukaryotic expression vector, and the recombinant eukaryotic expression vector can simultaneously express P-domain of nervous necrosis virus capsid protein and superfamily structural domain of garrupa interferon regulatory factor 3 protein. The bicistronic DNA vaccine for resisting the nervous necrosis virus, prepared by the invention, is transfected into a fish body, so that a nervous necrosis virus capsid protein P-domain and a superfamily structural domain of interferon regulatory factor 3 protein of a host can be well expressed. The prepared nervous necrosis virus resistant bicistronic DNA vaccine enhances the nervous necrosis virus infection resistance of fish bodies, reduces the death rate of the fish bodies infected with viruses, enhances humoral immunity of the fish bodies caused by the vaccine, causes all-around immune response of the fish bodies, and provides a good immune protection effect for the fish bodies.

The invention relates to construction and application of an IRES (Internal Ribosome Entry Site) mediated four GH (Growth Hormone) subtypes and IGF-I (Insulin-like Growth Factor I) bicistronic eukaryon co-expression vector. The construction comprises the following steps of: obtaining coding sequence design primers of four GH subtypes and the IGF-I by the comparison of the mRNA sequence and the protein sequence of the GH and the IGF-I; obtaining a target DNA of the four GH subtypes and the IGF-I through a PCR (Polymerase Chain Reaction) method by taking the DNA of the GH and the IGF-I in a laboratory as a template; connecting a PCR product to a pBS-T vector to obtain TA vectors of the GH and the IGF-I; carrying out XhO I and EcoR I double enzyme digestion on the TA vector and directionally inserting genes of the four GH subtypes into the pCI-IRES to successfully construct recombinant plasmids of the four GH subtypes: pCI-22-IRES, pCI-20-IRES, pCI-17-IRES and pCI-5-IRES; and carrying out Sam I and Not I double enzyme digestion on the TA vector of the IGF-I and respectively and directionally inserting genes of the IGF-I into such recombinant plasmids to successfully construct the IRES mediated four human GH subtypes and IGF-I bicistronic eukaryon co-expression vector: pCI-22-IRES-IGF-I, pCI-20-IRES-IGF-I, pCI-17-IRES-IGF-I and pCI-5-IRES-IGF-I. The invention provides convenient and efficient basic data for further studying biological characteristics and functions of the four GH subtypes and the IGF-I gene.

The invention discloses a primer group for detecting the reovirus and bicistronic virus of scylla serrata, comprising a reovirus forward primer ReoF, a reovirus reverse primer ReoR, a bicistronic virus forward primer DicF and a bicistronic virus reverse primer DicR, wherein each primer is specifically as follows: ReoF: 5-ACTCATAGAGCAGTCATGGG-3; ReoR: 5-ATATCGTCAGAATGTCGTTC-3; DicF: 5-GGATACTATGGATGATGTTTC-3; and DicR: 5-ACAAAATACCAGATAAAGCAA-3. The invention further discloses a kit containing the primer group and a detection method. The primer group, the kit and the detection method are shortin detection time, strong in specificity, high in detection sensitivity, and capable of simultaneously detecting the reovirus and bicistronic virus of scylla serrata.