108 results about "Cistron" patented technology

The present invention provides for transposon vectors encoding expression control region-traps and gene-traps. Also provided are dicistronic vectors. Certain embodiments of the invention contain internal ribosome entry sites.

The invention provides a DNA molecule comprising a multicistronic transcription unit coding for i) a polypeptide of interest, and for ii) a selectable marker polypeptide functional in a eukaryotic host cell, wherein the polypeptide of interest has a translation initiation sequence separate from that of the selectable marker polypeptide, and wherein the coding sequence for the polypeptide of interest is upstream from the coding sequence for the selectable marker polypeptide in said multicistronic transcription unit, and wherein an internal ribosome entry site (IRES) is present downstream from the coding sequence for the polypeptide of interest and upstream from the coding sequence for the selectable marker polypeptide, and wherein the nucleic acid sequence coding for the selectable marker polypeptide in the coding strand comprises a GTG or a TTG startcodon. The invention also provides methods for obtaining host cells expressing a polypeptide of interest, said host cells comprising the DNA molecules of the invention. The invention further provides the production of polypeptides of interest, comprising culturing host cells comprising the DNA molecules according to the invention.

Embodiments of the present invention relate to multicistronic vectors and methods for their design. Methods and compositions of the invention include a vector including at least two cistrons, wherein a first cistron includes a first promoter and a first nucleic acid sequence encoding one or more therapeutic agents, and wherein a second cistron comprises a second promoter and a second nucleic acid sequence encoding one or more RNA molecules that interfere with the expression of a biological response modifier or the therapeutic agent, wherein the expression of the first sequence is under control of the first promoter and expression of the second sequence is under control of the second promoter.

The 28-kDa outer membrane proteins (P28) of Ehrlichia chaffeensis are encoded by a multigene family consisting of 21 members located in a 23-kb DNA fragment in the genome of E. chaffeensis. Fifteen of these proteins are claimed herein as novel sequences. The amino acid sequence identity of the various P28 proteins was 20-83%. Six of 10 tested p28 genes were actively transcribed in cell culture grown E. chaffeensis. RT-PCR also indicated that each of the p28 genes was monocistronic. These results suggest that the p28 genes are active genes and encode polymorphic forms of the P28 proteins. The P28s were also divergent among different isolates of E. chaffeensis. The large repertoire of the p28 genes in a single ehrlichial organism and antigenic diversity of the P28 among the isolates of E. chaffeensis suggest that the P28s may be involved in immune avoidance.

Nucleic acids, including DNA constructs and RNA transcripts, capable of inducing coordinate expression of two to three cistrons upon direct introduction into animal tissues, are bi- or tri-cistronic polynucleotides of this invention include those encoding and co-expressing HIV gene products, genes encoding antigens unrelated to HIV, and immunostimulatory gene products, including but not limited to GM-CSF, interleukins, interferon and members of the B7 family of proteins which act as T-cell costimulatory elements. The methods and polynucleotides of this invention are generally applicable to co-ordinate expression in vivo of any two or more genes in a single cell.

The present invention is related to an antibiotic inducible / repressible genetic construct for controlling the transcription of a gene of interes: by a cell. The genetic construct comprises a bi-directional antibiotic controlled activator-responsive promoter / operator sequence which is located between the gene of interest and a cistron encoding a reverse antibiotic controlled transactivator and controls the transcription of the gene of interest and of the cistron.

The invention relates to polycistronic expression in gram-positive bacterium and in particular concerns polycistronic expression units comprising one or more gene endogenous to the gram-positive bacterium transcriptionally coupled to one or more genes exogenous to the bacterium.

Recombinant negative-strand viral RNA templates are described which may be used with purified RNA-directed RNA polymerase complex to express heterologous gene products in appropriate host cells and / or to rescue the heterologous gene in virus particles. The RNA templates are prepared by transcription of appropriate DNA sequences with a DNA-directed RNA polymerase. The resulting RNA templates are of the negative-polarity and contain appropriate terminal sequences which enable the viral RNA-synthesizing apparatus to recognize the template. Bicistronic mRNAs can be constructed to permit internal initiation of translation of viral sequences and allow for the expression of foreign protein coding sequences from the regular terminal initiation site, or vice versa.

The invention provides a method for detecting whether Araliaceae plant components exist in a sample and whether the sample is adulterate. The method for detecting whether Araliaceae plant components exist in a sample comprises the following steps: extracting total DNA (deoxyribonucleic acid) from a sample to be detected; by using the extracted total DNA as a template, carrying out PCR (polymerase chain reaction) amplification by using a primer pair, wherein the primer pair comprises SEQ ID NO:1 and SEQ ID NO:2; and carrying out analytical judgment on the PCR amplification product to identify whether Araliaceae plant components exist in the detected sample. The SEQ ID NO:1 and SEQ ID NO:2 are designed according to the ITS2 cistron sequence on an eucaryote rDNA (complementary deoxyribonucleic acid). Compared with the ITS2 universal sequence, the SEQ ID NO:1 and SEQ ID NO:2 can be used for carrying out amplification on the ITS 2 segment sequence of Araliaceae plants and can not be used for amplifying plant drugs of other families. The method can be used for detecting whether Araliaceae plant components exist in the sample, especially ginseng, pseudo-ginseng and / or American ginseng components.

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 provides a DNA molecule comprising a multicistronic transcription unit coding for i) a polypeptide of interest, and for ii) a selectable marker polypeptide functional in a eukaryotic host cell, wherein the polypeptide of interest has a translation initiation sequence separate from that of the selectable marker polypeptide, and wherein the coding sequence for the polypeptide of interest is upstream from the coding sequence for the selectable marker polypeptide in said multicistronic transcription unit, and wherein an internal ribosome entry site (IRES) is present downstream from the coding sequence for the polypeptide of interest and upstream from the coding sequence for the selectable marker polypeptide, and wherein the nucleic acid sequence coding for the selectable marker polypeptide in the coding strand comprises a GTG or a TTG startcodon. The invention also provides methods for obtaining host cells expressing a polypeptide of interest, said host cells comprising the DNA molecules of the invention. The invention further provides the production of polypeptides of interest, comprising culturing host cells comprising the DNA molecules according to the invention.

The polynucleotide construct of (1) or (2) below is used to perform ribosome display, CIS display and / or mRNA display in order to screen a Fab against an antigen of interest: (1) a polynucleotide construct which monocistronically comprises a ribosome-binding sequence, Fab first chain-coding sequence, linker peptide-coding sequence, Fab second chain-coding sequence and scaffold-coding sequence in this order, and further comprises at its 3′-end a structure necessary for maintaining a complex with the Fab encoded by itself, and (2) a polynucleotide construct which comprises a Fab first chain-expressing cistron and a Fab second chain-expressing cistron each containing a ribosome-binding sequence, a Fab first chain-coding sequence or Fab second chain-coding sequence, and a scaffold-coding sequence in this order, the first Fab-expressing cistron further comprising at its 3′-end a ribosome stall sequence, said Fab second chain-expressing cistron further comprising at its 3′-end a structure necessary for maintaining a complex with the Fab encoded by itself.

The invention provides a DNA molecule comprising a multicistronic transcription unit coding for i) a polypeptide of interest, and for ii) a selectable marker polypeptide functional in a eukaryotic host cell, wherein the polypeptide of interest has a translation initiation sequence separate from that of the selectable marker polypeptide, and wherein the coding sequence for the polypeptide of interest is upstream from the coding sequence for the selectable marker polypeptide in the multicistronic transcription unit, and wherein an internal ribosome entry site is present downstream from the coding sequence for the polypeptide of interest and upstream from the coding sequence for the selectable marker polypeptide, and wherein the nucleic acid sequence coding for the selectable marker polypeptide in the coding strand comprises a GTG or a TTG start codon. Also provided are methods for obtaining host cells expressing a polypeptide of interest, such host cells comprising DNA molecules of the invention. Further provided is the production of polypeptides of interest, comprising culturing host cells comprising the DNA molecules of the invention.

The invention provides a synthetic pyrethroid compound; the structure of the compound is shown by the formula A and comprises two cistron dextroisomers containing a formula B and a formula C; the molar ratio of the anti-form dextroisomer and the cis-form dextroisomer is 0.1:1-10:1, wherein R1 and R2 are halogen, halohydrocarbon, aliphatic hydrocarbon, aliphatic ether, alkoxy or hydrogen atom; X is aliphatic hydrocarbon of 1-3 carbon atoms, aliphatic ether, alkoxy, alkyne or nitrile grouping; compared with the prior art, the synthetic pyrethroid compound has high insecticidal activity; the invention further provides a preparation method of the synthetic pyrethroid compound and the application of preventing sanitary insect pests.

The present inventors conducted dedicated studies and successfully constructed expression vectors that enable high-level production of foreign gene-derived proteins in mammalian host cells, which comprise a translation-impaired dihydrofolate reductase gene cistron whose expression has been attenuated by altering the codons to the least frequently used codons in mammals; and a gene cassette which has a cloning site for incorporation of a foreign gene between a highly transcriptionally active promoter and a highly stable polyadenylation signal.

The present invention is directed generally to compositions and methods for expressing recombinant proteins in a mammalian host cell using a co-expressed transcriptional activator. In particular, the invention provides vectors, host cells, and methods of expressing at least one desired polypeptide by transfecting a mammalian host cell with cistrons encoding a transactivator, a desired polypeptide, and an apoptosis-protective protein.

The present invention is based on the discovery that a single lentiviral vector expressing multiple individual transcription factor proteins from a single multi-cistronic mRNA can reprogram a fibroblast cell to a stem cell-like cell. These reprogrammed induced pluripotent stem (iPS) cells are pluripotent. Additions of the Cre-LoxP sequences into the single lentiviral vector facilitate excision of the vector after reprogramming in achieved. Addition of a maker gene into the single lentiviral vector facilitates detection of the presence of the vector in an iPS. The invention provides compositions and methods of producing iPS cells using a single multi-cistronic lentiviral vector.

The invention provides a DNA molecule comprising a multicistronic transcription unit coding for i) a polypeptide of interest, and for ii) a selectable marker polypeptide functional in a eukaryotic host cell, wherein the polypeptide of interest has a translation initiation sequence separate from that of the selectable marker polypeptide, and wherein the coding sequence for the polypeptide of interest is upstream from the coding sequence for the selectable marker polypeptide in the multicistronic transcription unit, and wherein an internal ribosome entry site is present downstream from the coding sequence for the polypeptide of interest and upstream from the coding sequence for the selectable marker polypeptide, and wherein the nucleic acid sequence coding for the selectable marker polypeptide in the coding strand comprises a GTG or a TTG start codon. Also provided are methods for obtaining host cells expressing a polypeptide of interest, such host cells comprising DNA molecules of the invention. Further provided is the production of polypeptides of interest, comprising culturing host cells comprising the DNA molecules of the invention.

Nucleic acids, including DNA constructs and RNA transcripts, capable of inducing coordinate expression of two to three cistrons upon direct introduction into animal tissues, are. bi- or tri-cistronic polynucleotides of this invention include those encoding and co-expressing HIV gene products, genes encoding antigens unrelated to HIV, and immunostimulatory gene products, including but not limited to GM-CSF, interleukins, interferon and members of the B7 family of proteins which act as T-cell costimulatory elements. The methods and polynucleotides of this invention are generally applicable to co-ordinate expression in vivo of any two or more genes in a single cell.

The invention discloses a making method of antiepileptic drug alpha-asaryl, which comprises the follwoig steps: dissolving cistron-asaryl in the solvent; adding certain quantity of palladium compound as catalyst; stirring under indoor temperature or heating condition; filtering catalyst; decompressing; condensing the reacting liquid; recrystallizing residual through ligroin or skellysolve B to obtain the product.

The present inventors successfully constructed expression vectors that enable high-level production of foreign gene-derived proteins in mammalian host cells, which comprise a translation-impaired drug resistance gene cistron whose expression has been attenuated by altering the codons to the least frequently used codons in mammals; and a gene cassette which has a cloning site for incorporation of a foreign gene between a highly transcriptionally active promoter and a highly stable polyadenylation signal.

Recombinant NK cells, and especially recombinant NK-92 cells express a chimeric antigen receptor (CAR) having an intracellular domain of FcϵRIγ. Notably, CAR constructs with an intracellular domain of FcϵRIγ had a substantially prolonged duration of expression and significantly extended cytotoxicity over time. The CAR may be expressed from RNA and DNA, preferably as a tricistronic construct that further encodes CD16 and a cytokine to confer autocrine growth support. Advantageously, such constructs also enable high levels of transfection and expression of the recombinant proteins and provide a convenient selection marker to facilitate rapid production of recombinant NK / NK-92 cells.

The invention provides a method for evaluating the unintended effect of the molecular level of a transgenic plant based on a yeast two hybrid technology. The method comprises the following steps of: constructing an extraneous target protein as a bait fusion protein; constructing a cDNA (complementary Deoxyribonucleic Acid) library of a transgenic plant receptor parent as a prey fusion protein; and screening a prey fusion protein library with the prey fusion protein by adopting a yeast two hybrid method to obtain a positive cistron interacting with the extraneous target protein. According to the method, evaluation of the unintended effect of the molecular level of the transgenic plant is realized.

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 invention discloses a three-cistron expression vector, a preparation method and application and belongs to the technical field of gene engineering.The three-cistron expression vector comprises a nuclear matrix combining region sequence and a three-cistron sequence which is formed in the mode that two internal ribosomes enter a site sequence and connected; the structure of the three-cistron sequence is promoter-light chain signal peptide SPL-IRESwt-heavy chain signal peptide SPH-IRESatt-selection marker-Poly A.The vector can express a light chain, a heavy chain and a selection marker gene of an antibody at the same time, the problem that expression of a heavy chain, a light chain and a selection marker gene of a traditional vector is not balanced is solved, antibody quality is improved, and the cloned screening rate of positive cells is raised; meanwhile the nuclear matrix combining region sequence contained on the vector can further overcome transgene silencing, efficient and long-term expression of transgenes in host cells is achieved, on one hand, the antibody protein expression level is improved, and on the other hand, efficiency of screening of subsequent monoclonal antibody cell lines is improved.

The present invention relates to a process of obtaining recombinant lambdoid bacteriophage with high density display of functional peptides and proteins on surface of said phage comprising of: constructing a donor plasmid having a nucleotide sequence that defines the elements for replication of the vector in bacteria, a selectable marker, a nucleotide sequence flanked by two non-compatible recombination sequences, and an inducible cistron for expression of a capsid protein and a fusion protein; constructing a recipient phage having a nucleotide sequence that defines the lambdoid elements for replication and packaging of the vector into an assembled bacteriophage and encodes an inducible cistron for expression of a selectable marker flanked by two non-compatible recombination sequences; transferring the said donor plasmid to said recipient plasmid to obtain cointegrates; growing said cointegrates in selective liquid medium; harvesting phages displaying protein encoded by the foreign DNA encapsulated in said harvested phage particle.