35 results about "Vpr Protein" patented technology

The invention discloses a bacillus licheniformis host bacterium BL10 with multiple gene deletion, the accession number of which is CCTCCNO:M2013400. The host bacterium derives from a bacillus licheniformis WX-02 which partially or completely has deletion of ten genes. The ten genes comprises eight protease genes (mpr encoding a metalloprotease; vpr encoding a serine protease; aprX encoding an intracellular serine protease; epr encoding a minimal extracellular protease; bpr encoding a bacillus peptidase F; wprA encoding a protease combined with cell walls; aprE encoding an extracellular alkaline serine protease; and bprA encoding a bacillus peptidase F) and two extracellular secretory protein genes (hag encoding flagellin; and amyL encoding alpha-amylase). The BL10 has completely no extracellular protease activity and can reduce the degradation effect of a protease on a target protein. When the target protein is expressed by the expression host, the expression level is higher, and the host bacterium benefits protein expression enhancement.

The invention discloses a bacillus subtilis recombinant strain as well as a preparation method and application thereof and belongs to the field of a biological technology. The recombinant strain disclosed by the invention is obtained through knocking out or inactivating ten protease genes in bacillus subtilis; and the ten protease genes comprise: nprE, nprB, aprE, mpr, bpf, epr, vpr, wprA, spollAC and srfAC. The ten protease genes are knocked out through a homologous recombination method, the obtained bacillus subtilis recombinant strain is rapid in growth, and the expression amount of heterologous proteins is high. Therefore, the recombinant strain can be used as a host of efficient expression of the heterologous proteins.

The invention discloses bacillus subtilis used for high-efficiency expression and high-density culture of pullulanase, belonging to the technical field of bioengineering. According to the bacillus subtilis, a CRISPR/Cas9 gene editing system is adopted, 6 genes including nprB, bpr, mpr, vpr, epr and wprA of bacillus subtilis WS5 are knocked out in sequence, and the 6 bacillus subtilis strains including WSH6, WSH7, WSH8, WSH9, WSH10 and WSH11 are obtained. Bacillus subtilis WS5 and the 6 strains in competence are prepared, pullulanase expression plasmids are transformed, 7 pullulanase recombinant bacteria are obtained, and shake flask screening and tank screening adopting a 3-L tank are carried out. When the host strain obtained after screening is WSH9, the activity of pullulanase is the highest, fermentation is carried out for 78h by adopting the 3-L stank, and the activity of pullulanase can achieve 2449.6U/mL, which belongs to the extracellular highest level of pullulanase reported sofar.

The invention relates to APS method for shared VP- ring in ring network with ATM technology, which comprises: sending and receiving the APS cell defined with I.630 as reference and contained alert information and node space code address to detect logic link group state, realizing said protective switch to logic business with millisecond-level response speed; wherein, the switch is realized by modifying route list; when some node takes APS to self-ring business, the business of this ring also be automatic protection switched to match ring. This invention has less damage to business and high utility for bandwidth, and can match with APS of SDH.

The invention provides inducible CRISPRon or CRISPRi mouse embryo stem cells and a preparation method thereof. The mouse embryo stem cells are A2Lox.Cre cells and can reversibly express dCas9-VPR fusion protein or dCas9-KRAB fusion protein. The invention further provides a method for regulating and controlling gene expression and a kit. According to the inducible CRISPRon or CRISPRi mouse embryo stem cells provided by the invention, the construction method is simple and convenient, and the stable positive rate is up to 90 percent or higher; the genomic sequence is not edited, and gene expression is directly activated or inhibited; expression of the fusion protein has inducibility and reversibility; controllability and diversity on targeted regulation and control on the gene are achieved.

The invention discloses a voice data searching method and system, belonging to the data retrieval field. The method comprises: receiving a search instruction, voice information and a fuzzy searching scope inputted by a user through operating a search key; determining search conditions according to received voice information and fuzzy search scope, the search conditions being the number of characters separated between a keyword determined by the voice information inputted by the user and a keyword determined by the fuzzy searching scope, and/or the voiceprint recognition (VPR) characteristics of the voice information; and searching for voice data according to determined search conditions, and finding voice data matching the search conditions. Accordingly, the method can rapidly position voice data to be searched by users, and enhance user experience, and is fast and convenient.

The present invention provides a composition of matter, comprising: DNA encoding a viral Vpx protein fused to DNA encoding a protein. In another embodiment of the present invention, there is provided a composition of matter, comprising: DNA encoding a viral Vpr protein fused to DNA encoding a protein. The present invention further provides DNA, vectors and methods for expressing a lentiviral pol gene in trans, independent of the lentiviral gag-pol. A gene transduction element is optionally delivered to a lentiviral vector according to the present invention. Also provided are various methods of delivering a virus inhibitory molecule to a target in an animal. Further provided is a pharmaceutical composition.

The invention relates to double bond small molecule disturbance ribonucleic acid to prevent and cure AIDS. The double bond small molecule disturbance ribonucleic acid is RNA double bond molecule, which is called siRNA double bond molecule that has 19 base pairings. Two protrude basic groups dT are on the ends of 5' of positive bond and negative bond. GC content is 40-55%. Target sequence is gag gene, pol gene, vif gene, vpr gene, env gene, and nef gene selected from HIV gene. The positive bond sequence of siRNA double bond molecule is selected from SEQ ID NO.1-116, and the negative bond is corresponding to positive sequence. The invention conquers the problem of validity decreasing or losing effectiveness of siRNA because of the mutation of HIV-I virus.

The present invention provides screening methods for identifying a compound that induces loss of the lentiviral protein Vpr; screening methods for identifying compounds that inhibit the peptidyl-prolyl cis/trans isomerase (PPIase) activity of a protein that catalyzes cis-trans isomerization of cis-peptidylprolyl bonds in Vpr; and compounds identified by the screening methods. The compounds are useful for treating a lentiviral infection. The present invention further provides methods of inducing loss of the lentiviral protein Vpr; methods of inhibiting lentivirus viral replication; and methods of treating a lentivirus infection in an individual. The methods generally involve administering to an individual infected with the lentivirus an effective amount of a compound that induces Vpr loss and/or that inhibits PPIase activity of a protein that catalyzes cis-trans isomerization of cis-peptidylprolyl bonds in Vpr.

This invention provides novel methods for identifying agents that inhibit HIV infection. The anti-HIV agents are identified by screening test compounds for ability to modulate a biological activity of isopeptidase T (IsoT), e.g., its isopeptidase activity or its binding to another molecule such as viral protein R (Vpr). Such IsoT modulators can be further examined for their activity in inhibiting an activity indicative of HIV infection or HIV replication. These novel anti-HIV agents are useful in the prevention or treatment of HIV infection and conditions associated with or caused by HIV infection.

The invention relates to the tumour genes treatment, as more specific, it relates to a new tumour therapeutic agent and its usage. The tumour therapeutic agent contains human immunodeficiency virus Vpr gene, which is in control of a promoter and express specificly in the tumor structure. The invention can be used in tumor treatment, and can be the intensifier in tumor chemical therapy or radiation therapy.

The invention relates to the technical field of biology, and particularly discloses a recombinant immunodeficiency plasmid and virus and application thereof. The immunodeficiency plasmid comprises LTR, gag gene, nucleotide sequence disclosed as SEQ ID No.1, vif gene, vpr gene, vpx gene, tat gene, rev gene, env gene and nef gene. The nucleotide sequence disclosed as SEQ ID NO.1 is introduced to the pathogenic SIVmac239 full-length plasmid frame to construct the immunodeficiency plasmid comprising a plurality of drug target spots, and packaging is performed in the 293T cell to generate immunodeficiency virus. The virus has the capacity for infecting target cells and the capacity of efficient replication, and is applicable to researching human immunodeficiency virus (HIV) in-vivo infection process, pathogeny characteristic, pathogenesis and immunoreaction and screening anti-HIV drugs and anti-HIV drug application strategies.

The invention discloses a method for extracting enhanced VPR (viral protein regulatory) protein and plasma free nucleic acid. An amino acid sequence of the enhanced VPR protein is obtained by the mutation of the amino acid sequence shown in SEQ ID NO:1; the mutation at least comprises one of following mutation sites: a 55th site, a 102nd site, a 134th site and a 166th site; the mutation of tyrosine at the 55th site is phenylalanine; the mutation of serine at the 102nd site is threonine; the mutation of serine at the 134th site is isoleucine; the mutation of serine at the 166th site is threonine; or, the amino acid sequence of the enhanced VPR protein has the mutation sites in the amino acid sequence with mutation, and has more than 98% homology with the amino acid sequence with mutation. The enhanced VPR protein has the advantage that the activity is higher at normal temperature, so that the heating is not required in the plasma free nucleic acid extraction process, and the degradationof nucleic acid is reduced.

The invention relates to the field of genetic engineering, in particular to a fusion immunotoxin B-L-Vpr gene as well as cloning and heterologous expression. The gene has a base sequence in Sequence Table SEQ ID No:1, meanwhile, protein encoded by the fusion immunotoxin B-L-Vpr gene has an amino acid sequence in SEQ ID No:2. In the invention, a single-chain antibody for resisting HER-2 is regarded as a targeted molecule and Vpr protein is regarded as an effector molecule. The targeted molecule and the effector molecule form the fusion immunotoxin B-L-Vpr through the connection of small peptides and effectively express the fusion immunotoxin protein in colon bacillus. The fusion immunotoxin can have a specific inhibitive effect on a tumor cell over-expressed by a HER-2 receptor.

Novel packaging cell lines which produce recombinant retrovirus, free of detectable helper-virus are disclosed. Also disclosed are methods of making the cell lines and methods of producing recombinant retroviruses from the cell lines. Retroviruses produced by the cell lines include lentiviruses, such as HIV, capable of transfering heterologous DNA to a wide range of non-dividing cells. The packaging cells contain at least three vectors which collectively encode retroviral gag, pol, and env proteins, wherein the gag and pol genes are separated, in part, onto two or more different vectors. This is made possible by fusing Vpr or Vpx to pol proteins separated from gag so that the proteins are targeted to assembling virions. Among other advantages, the packaging cells provide the benefit of increased safety when used in human gene therapy by virtually eliminating the possibility of molecular recombination leading to production of replication-competant helper virus.

Macrophage colony-stimulating factor (M-CSF) is important for human immunodeficiency virus-type 1 (HIV-1) infection, replication and survival of infected cells. The mechanism(s) by which HIV-1 infection increases M-CSF production are, however, poorly understood. Here, we report that HIV-1 Vpr enhances M-CSF promoter activity and production in primary human monocytes and macrophages. Vpr activates M-CSF transcription through four C/EBP beta binding sites present within the M-CSF promoter, possibly through increased phosphorylation of C/EBP beta. RU486 (mifepristone) blocked Vpr-mediated up-regulation of M-CSF, suggesting that Vpr activates M-CSF promoter activity via the glucocorticoid pathway. The invention provides new avenues for therapeutic interventions in HIV-1 infection and other diseases involving M-CSF dysregulation (including malignancy, osteoporosis, autoimmune disorders, arthritis, and obesity) using glucocorticoid antagonists and modulators of C/EBP beta activity.

The invention relates to fusion proteins comprising the amino acid sequence of at least four HIV proteins selected from Vif, Vpr, Vpu, Rev, Tat and Nef or derivatives of the amino acid sequence of one or more of said proteins, wherein the fusion protein is not processed to individual HIV proteins having the natural N and C termini. The invention further concerns nucleic acids encoding said proteins, vectors comprising said nucleic acids, and methods for producing said proteins. The fusion protein, nucleic acids and vectors are usable as vaccines for the at least partial prophylaxis against HIV infections.

The invention discloses application of a Vpr protein N-terminal amino acid polypeptide for regulating and controlling tumor cell apoptosis in preparation of antitumor drugs. The polypeptide is derived from 23rd-37th amino acids at the N terminal of HIV protein Vpr. And the sequences of the 23rd-37th amino acids at the N end of the Vpr protein are SEQ ID NO. 1. The 23rd-37th amino acids at the N end of the Vpr protein and an adenine nucleotide transporter 1 or an adenine nucleotide transporter 3 act together to promote apoptosis of tumor cells; and the 23rd-37th amino acids complex with adenine nucleotide transporter 2 to inhibit tumor cell apoptosis. The amino acid polypeptide sequence is derived from virus protein, and the reagent can regulate and control proliferation and apoptosis of tumor cells through a new mechanism and can be used for biological treatment of tumors.

The HIV-1 accessory gene vpr encodes a conserved 96-amino acid protein that is necessary and sufficient for the HIV-1-induced block of cellular proliferation and induction of apoptosis. Expression of vpr in CD4⁺ lymphocytes results in G2 arrest, followed by apoptosis. ATR, as a cellular factor that mediates Vpr-induced cell cycle arrest, is required for activation of the Breast Cancer-Associated Protein-1 (BRCA1). In addition, the Growth Arrest and DNA Damage protein (GADD45) is upregulated by Vpr in an ATR-dependent manner. Posttranscriptional silencing of either ATR or GADD45 leads to nearly complete suppression of the pro-apoptotic and/or cell cycle arrest effect of Vpr.

Conjugated compositions comprising a fragment of HIV-1 Vpr or a non-HIV-1 Vpr protein conjugated to a therapeutic compounds and methods of using the same to deliver therapeutic compounds to a cell's nucleus or for the preparation of drug delivery particles are disclosed. Functional fragments of HIV-1 Vpr and functional non-HIV-1 Vpr proteins, and pharmaceutical compositions comprising the same are disclosed. Methods of inhibiting cell proliferation and methods of treating an individual who has a hyperproliferative disease are disclosed. Methods of identifying compounds that inhibit Vpr protein binding to the p6 domain of p55 or to p6 protein and kits for performing such methods are disclosed.