40 results about "RNA polymerase II" patented technology

The present invention relates to polyamides capable of inhibiting ARE-, GRE- and ERE-mediated gene regulation in cells. The present invention also relates to polyamides capable of modulating the activity of RNA polymerase II and p53. The invention also relates to methods to treat diseases related to ARE-, GRE- and ERE-mediated gene regulation and to RNA polymerase II and p53 activity.

The invention relates to the field of recombinant DNA technology and, in particular, the development of vectors for the expression of recombinant proteins. Expression of heterologous genes in eukaryotic cells requires transcription by RNA polymerase II, which is driven by c / s-acting genetic elements known as promoters and enhancers. The invention provides promoters derived from the guinea pig cytomegalovirus early- immediate promoter / enhancer, useful for obtaining high levels of expression of recombinant proteins. Also disclosed are eukaryotic expression vectors comprising such promoters, which are capable of providing increased levels of expression over that obtainable from human or murine cytomegalovirus enhancer / promoter elements in many cell types.

The invention provides a use of RNA polymerase II subunit A C-terminal domain phosphatase (CTDP1) or a fragment thereof in preparation of a reagent used for diagnosing Behcet disease (BD). The positive rate of the CTDP1 or the fragment thereof in a BD patient is remarkably higher than that of normal control, a Western blot verification result displays that an anti-CTDP1 antibody can be detected in the serum of the BD patient, which proves that the CTDP1 or the fragment thereof can be used for a detection marker of BD.

Provided is a method of modulating the expression of a gene containing expanded nucleotide repeats in a cell, comprising: inhibiting the biological activity of SPT4 or SUPT4H; and regulating the formation of R-loops. The inhibition step can effectively reduce the expression of the gene containing the expanded nucleotide repeats and the regulatory step can further enhance the inhibition step. The inhibition step and the regulation step are for the purpose of regulating gene expression by interfering the capacity of RNA polymerase II transcribing over a DNA template with lengthy nucleotide repeats.

The invention discloses an empty carrier for construction of a siRNA expression vector, and a recombinant vector for siRNA and application thereof. The empty carrier, from terminal 5' to terminal 3', comprises: 1) a CMV-derived RNA polymerase II dependent promoter sequence; 2) a pri-MiR21 front fragment sequence as represented by SEQ ID No. 1; 3) the sequence of a cleavage site of restriction endonuclease; 4) a pri-MiR21 rear fragment sequence as represented by SEQ ID No. 2; and 5) a terminator sequence. The recombinant vector for siRNA is obtained by inserting the sequence of siRNA sense sequence-MiR21 ring structure sequence-siRNA antisense sequence to the cleavage site of the restriction endonuclease. According to the invention, since an RNA Pol II promoter is employed, shRNA can be conveniently and effectively connected to the skeleton of pri-MiR21 and be stably expressed through the RNA Pol II promoter, high efficiency expression is realized, and efficiency of knockout and deletion of RNAi is improved.

The invention relates to the field of biotechnology and aims to provide a method for constructing a molecularly marked CSFV (classical swine fever virus) attenuated vaccine. The method comprises following steps: a recombinant plasmid pA-F123 containing genome 5' half-length of a CSFV vaccine strain C and a recombinant plasmid pE-B-Erns containing Erns genes of a BVDV (bovine viral diarrhea virus) strain VEDEVAC are taken as templates, 20bp of homologous fragments are designed at two ends, and by means of a one-step orient cloning kit, a recombinant plasmid pA-B-Erns-F123 is obtained; an F456 fragment containing genome 3' half-length of the CSFV vaccine strain C is cut off from a recombinant plasmid pB-F456 through BamH I and Sal I and cloned into pA-B-Erns-F123 under the action of the same enzyme, and a recombinant plasmid pA-B-Erns-FL is obtained. By virtue of establishment of a reverse genetic manipulation technology, a new way is developed for research of CSFV gene functions and novel vaccines. An exogenous tag is inserted into a viral genome with the reverse genetic manipulation technology for research of virus replication, virus encoded protein functions and anti-virus drug screening, and an RNA (ribonucleic acid) polymerase II system can efficiently and stably save CSFV infectious cDNA (complementary deoxyribonucleic acid ) cloning.

An isolated or a purified nucleic acid sequence for enhancing expression levels of a protein of interest, in 5′ to 3′ direction, comprises: a cytomegalovirus (CMV) promoter; an eEF-1 intron repeat (eEF-1 IR); and a regular sequence, which comprises: at least one tag element, a fixable linker sequence, wherein the fixable sequence is TEV sequence; and a multiple cloning site (MCS). By means of the array of the specific promoter and eEF-1 IR, the expression and purity of the recombinant protein could be enhanced; wherein eEF-1 IR can reduce the length of vector and assist RNA polymerase II transcription. Besides, TEV sequence of the fixable linker sequence of the regular sequence can remove a tag on recombinant protein; a specific target can be inserted into the multiple cloning site.