40 results about "Start site" patented technology

The invention provides antisense antiviral compounds and methods of their use and production in inhibition of growth of viruses of the Filoviridae family, and in the treatment of a viral infection. The compounds and methods relate to the treatment of viral infections in mammals including primates by Ebola and Marburg viruses. The antisense antiviral compounds are morpholino oligonucleotides having: a) a nuclease resistant backbone, b) 15-40 nucleotide bases, and c) a targeting sequence of at least 15 bases in length that hybridizes to a target region selected from the following: i) the AUG start site region of VP35, as exemplified by SEQ ID NOS:67-71 or ii) the AUG start site region of VP24, as exemplified by SEQ ID NOS:72-76.

The present invention provides antisense antiviral compounds, compositions, and methods of their use and production, mainly for inhibiting the replication of viruses of the Filoviridae family, including Ebola and Marburg viruses. The compounds, compositions, and methods also relate to the treatment of viral infections in mammals including primates by Ebola and Marburg viruses. The antisense antiviral compounds include phosphorodiamidate morpholino oligonucleotides (PMOplus) having a nuclease resistant backbone, about 15-40 nucleotide bases, at least two but typically no more than half piperazine-containing intersubunit linkages, and a targeting sequence that is targeted against the AUG start site region of Ebola virus VP35, Ebola virus VP24, Marburg virus VP24, or Marburg virus NP, including combinations and mixtures thereof.

The invention belongs to the technical field of genomics, and particularly relate to a method for simplifying CAGE-seq library establishing.The method comprises the steps that mRNA and lncRNA containing cap structures are enriched, a 5'-termnal connector is specifically added, after fragmentation processing, reverse transcription is carried out, and 5'-terminal information is further enriched.According to the whole method, an existing mature RNA-seq kit of the gnomegen company is mixed.The method is easier to operate and high in success rate, the 5'-terminal information can be specifically enriched, the method is more helpful for verifying the transcriptional start site of mRNA in a whole cell, and thus acquiring accurate promoter information and 5'UTR information.The method is a powerful mode for studying gene expressions and is suitable for gene regulatory network studies.

The invention relates to the field of molecular biology medicine. saRNA molecules of TPO genes are synthesized, the sequence of the saRNA molecules and the sequence SEQ ID NO:1 of the upstream 6,000bp and downstream 1,000bp areas of a TPO gene transcriptional start site are complementary, and low-expression or non-expression TPO genes in tumor cells can be effectively activated. A biological effect of promoting the tumor cells to hold radionuclide 131I is achieved, a novel method and novel molecular targets are provided for combining the gene technology with the radionuclide therapy for targeted therapy of tumors.

The invention relates to a controllable genome-modified plasmodium, a recombinant expression vector and the construction method and application of the controllable genome-modified plasmodium and the recombinant expression vector. The recombinant expression vector comprises a gene targeting long homologous arm, a gene targeting short homologous arm, a tetracycline repression protein gene expression cassette, a pyrimethamine resistance gene expression cassette and a target gene expression cassette, wherein the tetracycline repression protein gene expression cassette, the pyrimethamine resistance gene expression cassette and the target gene expression cassette are located between the gene targeting long homologous arm and the gene targeting short homologous arm, and tetracycline operator gene sequences are inserted in multiple transcriptional start sites of a target gene promoter, so that the recombinant expression vector can be used for conditional research of the functions of a certain functional gene in a plasmodium genome. Furthermore, a functional gene expression sequence, corresponding to a target gene, in the plasmodium genome is knocked out by means of the gene knockout technique; meanwhile, the recombinant expression vector is transfected into a plasmodium with genes knocked out, so that the controllable genome-modified plasmodium is obtained; a new technical scheme is provided for further research of the functions of all functional genes in the plasmodium genome, and application prospects are broad.

The serial double function cos carriers, including pJTU390, pJTU391, pJTU407, pJTU408, pJTU409, pJTU410, pJTU411 and pJTU412, suitable for streptomycede chromosome gene knock-out feature that each of them consists of following DNA elements: colibacillus plasmid Coie1 duplicating initiation site ori, ampicilin resistance gene bla, streptomycete plasmid pIJ101 duplicating initiation site and duplicon, thiactin resistance gene tsr, lambda bacteriophage cos site, RP4 conjugal transfer start site oriT, promoter capable of being identified specifically by T3 and T7 bacteriophage RNA polymerase, and neomycin resistance gene neo from Tn5 and containing no promoter. The present invention contains colibacillus plasmid, streptomycete plasmid duplicon and resistance screening mark simultaneously, has functions of autonomous duplicating and inheriting simultaneously in two kinds of hosts, and facilitates in vitro genetic operation and gene function analysis.

The present invention relates to expression vectors for the heterologous expression of a nucleic acid sequence of interest in mammalian cells, the vectors comprising a chimeric promoter regulatory sequence being operably linked to a nucleic acid sequence to be expressed, wherein the chimeric promoter regulatory sequence comprises a cytomegalovirus promoter sequence derived from murine cytomegalovirus or from human cytomegalovirus and being operably linked to the transcriptional start site of the nucleic acid sequence to be expressed; and a cytomegalovirus upstream region and / or enhancer sequence derived from human and / or the simian cytomegalovirus, wherein the upstream region and / or enhancer sequence is located 5′ of and operably linked to the murine or the human promoter sequence, and wherein the chimeric promoter regulatory sequence comprises sequence elements being derived from at least two of the group consisting of murine cytomegalovirus, human cytomegalovirus and simian cytomegalovirus. In particular embodiments, the chimeric promoter regulatory sequence comprises sequence elements derived from the murine or the human cytomegalovirus IE1 promoter and from the human and / or the simian cytomegalovirus IE1 region. The invention also relates to mammalian host cells transfected with such expression vectors, a method for heterologous expression of a nucleic acid sequence in a mammalian host cell by employing such expression vectors, and the use of such expression vectors for the heterologous expression of a nucleic acid sequence.

Provided herein are antisense oligonucleotides, compositions comprising antisense oligonucleotides, and methods for the use of antisense oligonucleotides in manipulating translation. Expression of isoforms of proteins expressed from different start codons of the same transcript are inhibited by antisense oligonucleotides, which may also enhance expression of non-target isoforms.

Staphylococcus aureus is notorious for developing resistance to virtually all antibiotics to which it is exposed. Staphylococcal phage 2638A endolysin is a peptidoglycan hydrolase that is lytic for S. aureus when exposed externally, making it a new antimicrobial candidate. It shares a common protein organization with over 40 other staphylococcal peptidoglycan hydrolases: a CHAP endopeptidase domain, a mid-protein amidase 2 domain and a C-terminal SH3b cell wall binding domain. It is the first phage endolysin reported with a cryptic translational start site between the CHAP and amidase domains. Deletion analysis indicates that the amidase domain confers most of the lytic activity and requires the full SH3b domain for maximal activity. It is common for one domain to demonstrate dominant activity over another; however, the phage 2638A endolysin is the first to show high amidase domain activity dominant over the N-terminal CHAP domain, an important finding for targeting novel peptidoglycan bonds.

The invention relates to a controllable genome-modified plasmodium, a recombinant expression vector and the construction method and application of the controllable genome-modified plasmodium and the recombinant expression vector. The recombinant expression vector comprises a gene targeting long homologous arm, a gene targeting short homologous arm, a tetracycline repression protein gene expression cassette, a pyrimethamine resistance gene expression cassette and a target gene expression cassette, wherein the tetracycline repression protein gene expression cassette, the pyrimethamine resistance gene expression cassette and the target gene expression cassette are located between the gene targeting long homologous arm and the gene targeting short homologous arm, and tetracycline operator gene sequences are inserted in multiple transcriptional start sites of a target gene promoter, so that the recombinant expression vector can be used for conditional research of the functions of a certain functional gene in a plasmodium genome. Furthermore, a functional gene expression sequence, corresponding to a target gene, in the plasmodium genome is knocked out by means of the gene knockout technique; meanwhile, the recombinant expression vector is transfected into a plasmodium with genes knocked out, so that the controllable genome-modified plasmodium is obtained; a new technical scheme is provided for further research of the functions of all functional genes in the plasmodium genome, and application prospects are broad.

The invention provides a design method and application of a primer set. According to the needs of amplification length, the sequence before the FR1 start site of the antibody is subtly extracted from the upstream of the FR1 start site of the antibody, and the sequence is extracted from the 5' end of the primer set. Starting from the first base, the fixed-length primer sequences are sequentially shifted and cut to form a candidate primer library. After cluster analysis, the primer set of the immune group library is screened. The primer set obtained by this method can significantly improve the immune group of the multiplex PCR method. The coverage and experimental efficiency of library amplification, reducing the amplification mismatch rate, the method is simple, and the cost is saved.

Methods for screening a polynucleotide library for a polypeptide with a property of interest in a filamentous fungal host cell, in a manner which allows quick and easy subsequent characterization of the polypeptide, using an expression cloning vector comprising at least a polynucleotide encoding a selectable marker in which the translation initiation start site of the marker-encoding sequence comprises a crippled consensus Kozak sequence, a fungal replication initiation sequence, and a promoter with a cloning-site into which the library is cloned, and a transcription terminator.