58 results about "Provirus" patented technology

The present invention relates to methods and compositions for the elucidation of mammalian gene function. Specifically, the present invention relates to methods and compositions for improved mammalian complementation screening, functional inactivation of specific essential or non-essential mammalian genes, and identification of mammalian genes which are modulated in response to specific stimuli.In particular, the compositions of the present invention include, but are not limited to, replication-deficient retroviral vectors, libraries comprising such vectors, retroviral particles produced by such vectors in conjunction with retroviral packaging cell lines, integrated provirus sequences derived from the retroviral particles of the invention and circularized provirus sequences which have been excised from the integrated provirus sequences of the invention. The compositions of the present invention further include novel retroviral packaging cell lines.

A vector comprising an HIV segment and a heterologous gene segment, which produces a replication competent and an infective HIV virus is disclosed. When the heterologous gene is a marker gene, the spread of the virus can be observed in both in vitro and in vivo systems. The use of this vector in establishing methods for screening anti-viral compounds is also disclosed.

Improved molecular vaccines comprise nucleic acid vectors that encode a fusion polypeptide that includes polypeptide or peptide physically linked to an antigen. The linked polypeptide is one that (a) promotes processing of the expressed fusion polypeptide via the MHC class I pathway and/or (b) promotes development or activity of antigen presenting cells, primarily dendritic cells. These vaccines employ one of several types of nucleic acid vectors, each with its own relative advantages: naked DNA plasmids, self-replicating RNA replicons and suicidal DNA-based on viral RNA replicons. Administration of such a vaccine results in enhance immune responses, primarily those mediated by CD8+ cytotoxic T lymphocytes, directed against the immunizing antigen part of the fusion polypeptide. Such vaccines are useful against tumor antigens, viral antigens and antigens of other pathogenic microorganisms and can be used in the prevention or treatment of diseases that include cancer and infections.

Methods and compositions containing a phorbol ester or a derivative of a phorbol ester are provided for the treatment of cytopathic diseases. Cytopathic diseases may be caused by a variety means suchas viral infections like HIV and AIDS, or the development of neoplasms in a mammalian subject. The methods and compositions of the invention are effective for inhibiting de novo HIV infection, upregulating viral expression from latent provirus, inhibiting HIV-induced cytopathic effects, down regulating the HIV receptor, increasing ThI cytokine expression, decreasing Th2 cytokine expression, increasing ERK phosphorylation, inducing apoptosis in malignant cells, inducing remission, maintaining remission, as chemotherapeutic agents, as well as for decreasing symptoms of cytopathic diseases and opportunistic infections that may accompany such diseasesl. Additional compositions and methods are provided which employ a phorbol ester or derivative compound in combination with at least one additional agent such as those used in HAART protocols, therapeutic agents used to treat opportunistic infections due to HIV, or chemotherapeutic agents to yield more effective treatment tools against cytopathic diseases in mammalian subjects.

The development of general approaches for the isolation of efficient antivirals is becoming increasingly important. The genetic suppressor element (GSE) technology is an approach based on the functional expression and selection of efficient genetic inhibitors from random fragment libraries derived from a gene or genome of interest. We have applied this technology to isolate potent genetic inhibitors against the human immunodeficiency virus type 1 (HIV-1) The strategy employed involved the following steps: 1) fragmenting the HIV-1 genome into 100-700 base pair (bp) fragments; 2) inserting the fragments into expression vectors to form an expression library; 3) transferring the expression library into a population of cells (e.g., OM10.1) containing an inducible latent HIV-1 provirus; 4) selecting a subpopulation of cells which contain a subset of the expression library enriched for HIV-1 GSE by monitoring the expression of a cellular (e.g., CD4) or viral (e.g., p24) marker associated with HIV infection; 5) recovering the GSE from the selected cell population. The GSEs identified clustered in seven narrowly defined regions of the HIV-1 genome and were found to be functionally active. These elements are potential candidates for the gene therapy of AIDS. The developed approaches can be applied to other viral pathogens, as well as, for the identification of cellular genes supporting the HIV-1 life cycle.

The invention relates to the field of molecular biology and immunology, in particular to an application of an HBcAg (hepatitis B core antigen) virus-like particle serving as a cervical cancer therapeutic vaccine carrier. A preparation method comprises steps as follows: an HPV16 E749-57CTLs epitope peptide fragment is selected, a DNA (deoxyribose nucleic acid ) fragment of the HPV16 E749-57CTLs epitope peptide fragment is inserted between 78 and 79 amino acids of the HBcAg through genetic recombination, an obtained recombinant plasmid pHBcAg-E749-57 is converted into Escherichia coli DH5alpha, and an HBcAg virus-like particle vaccine presenting E749-57 is obtained after induction expression and purification. After a tumor-bearing mouse is immunized with the virus-like particle vaccine, the body of the mouse can be induced to generate a higher HPV16E7 specific cellular immunologic response, and growth of tumors is remarkably inhibited.

The invention provides a primer and a method for detecting HTLV (human T-lymphotropic virus)-I and HTLV-II proviruses. The primer comprises a specific primer and a probe for detecting an HTLV-I provirus and a specific primer and a probe for detecting an HTLV-II provirus, the two primers and the two probes are added in the same PCR (polymerase chain reaction) pipe according to a reasonable concentration ratio, and a sample is detected by means of optimized Real-time PCR reaction conditions. The detection method can be used for detecting whether HTLV infection exists before serological changes, thereby greatly shortening the window phase; meanwhile, compared with a conventional serological detection method, the detection method has the advantages of short detection period, high specificity, high accuracy, high sensitivity, little dependence on the conditions, low pollution risk and the like. The detection result is beneficial to monitor the change of the carrying capacity of the HTLV provirus of a patient and limit the propagation of the HTLV virus.

The invention discloses a trace nucleic acid releasing agent. The releasing agent realizes that hepatitis B virus nucleic acid extraction and fluorescent augmentation are directly performed in the same tube, radically solves the problems of nucleic acid loss and pollution caused by a multi-step nucleic acid extraction method, and is applicable to the fluorescent PCR detection of a whole blood sample for the first time; the releasing agent adopts quantificational provirus serum as a quantificational standard and realizes the whole process monitoring of the standard to the sample; the releasing agent adopts double pairs of forward primers and reverse primers and a probe decorated by forward fluorescence and reverse fluorescence to perform fluorescent PCR augmentation, further to improve the sensitivity and the augmentation efficiency of the releasing agent; blue promoting fluorescer is introduced to ensure that prepared PCR operating liquid is light blue, and has the functions of steady fluorescent signals, strengthened augmentation efficiency and so on, without affecting the fluorescent detection; the releasing agent sets two temperature gradient programs, adopts a rear-section fluorescence acquisition method, is convenient to analyze and improves the service life of an instrument; compared with the prior method, the technology has revolutionary innovation.

The present invention relates to the expression and screening of genomic DNA sequences encoding uncharacterized genes and proteins. The present invention provides systems utilizing unique features of retroviral replication to analyze uncharacterized genes derived from genomic DNA samples. In preferred embodiments, a segment of genomic DNA is inserted between 5′ and 3′ viral long terminal repeats (LTRs) in a vector (e.g., a plasmid, cosmid, or artificial chromosome vector). The resulting vector (or library of vectors containing a plurality of independent genomic sequences) is then introduced into a retroviral packaging cell. The resulting provirus or proteins expression from the provirus are then analyzed.

The present invention provides an approach for developing an algorithm for determining the effectiveness of anti-viral drugs based on a comprehensive analysis of paired phenotypic and genotypic data guided by phenotypic clinical cut-offs. In one aspect, the algorithm allows one to provide a patient with effective treatment. It helps predict whether an infected individual will respond to treatment with an anti-viral compound, thereby allowing an effective treatment regimen to be designed without subjecting the patient to unnecessary side effects. Also, by avoiding the administration of ineffective drugs, considerable time and money is saved.

The invention belongs to the field of gene therapy and relates to preparation and an application of an expression vector for targeted inhibition of HIV-1 provirus expression by use of embedded DNA methyltransferase adopting a zinc finger structure. The expression vector carrying the DNA methyltransferase adopting the zinc finger structure is proved to be able to inhibit silence of HIV-1 provirus expression in HIV-1 infection and latent infection cell models, the vector can effectively inhibit virus replication in virus infected cells and latent infected cells for 15-40 days or above. Besides,provirus expression silence caused by the expression vector carrying the embedded methyltransferase adopting the zinc finger structure can be inhered among cell generations and has certain safety in cells. A new idea is provided for curing AIDS.

The present invention is directed to a method for preparing an expression vector encoding a tailored recombinase, wherein said tailored recombinase recombines asymmetric target sites within the LTR of proviral DNA of a retrovirus inserted into the genome of a host cell and is useful as means for excising the provirus from the genome of the host cell. The present invention further relates to an in vitro-method of optimising the treatment of a retroviral infection of a subject and to the use of tailored recombinases for the preparation of pharmaceutical compositions for reducing the viral load in a subjected infected by a retrovirus.

The invention belongs to the field of veterinary products, and discloses a preparation method of duck tembusu virus infectious cDNA (complementary deoxyribonucleic acid), which comprises the following steps: step 1, constructing subgenome plasmids, the subgenome plasmids are plasmids pF1, pF2, pF3 and pF4, and the plasmids pF1, pF2, pF3 and pF4 respectively contain target fragments F1, F2, F3 and F4; 2, obtaining target fragments: obtaining the target fragments f1, f2, f3 and f4 for in-vitro connection from the plasmids pF1, pF2, pF3 and pF4; and step 3, connecting target fragments f1, f2, f3 and f4 in vitro to obtain the duck tembusu virus infectious cDNA. The method can successfully clone the recombinant virus which is completely the same as the original virus strain and has no mutation.

The present invention relates to methods and agents for preventing the establishment of HIV-1 latent reservoirs or for reducing the size of the reservoirs. Specifically, the disclosure provides methods and agents for preventing the establishment of HIV-1 latent reservoirs or for reducing the size of the reservoirs, the methods comprising administering to the subject a therapeutically effective amount of an isolated anti-HIV antibody, and administering to the subject two or more viral transcription inducers in effective amounts to induce transcription of an HIV provirus in the cells. Further provided are antibodies and viral transcription inducers used in the methods.

The invention relates to a coronavirus vaccine as well as a preparation method and an application thereof. The coronavirus vaccine comprises a nano-carrier and coronavirus coated in the nano-carrier,wherein the nano-carrier comprises chitosan and/or a chitosan derivative. The preparation method of the coronavirus vaccine comprises the following steps: mixing coronavirus with the carrier, and thenmixing with weak acid to form nanoparticles, namely the coronavirus vaccine. According to the invention, the coronaantigen virus is creatively coated in chitosan and/or the chitosan derivative to finish a sterilization procedure in a physical mode, so that compared with a traditional chemical sterilization mode, the production effect of the vaccine is improved, the production cost is saved, and the risk of other diseases caused by mutation and reversion of the coronaantigen virus is reduced; in addition, due to the biocompatibility and biodegradability of the chitosan or the derivative thereof, the vaccine has very good biological safety.

The invention relates to a quick detecting method of bovine leukemia virus, which directly takes blood as a detecting sample, does not need special treatment, and is convenient to collect, store and transport. Primers are designed to amplify LTR of bovine leukemia virus (BLV) provirus, which ensured the accuracy and stability of the method. Based on the nested PCR detecting method, the quick detecting method of bovine leukemia virus can detect bovine leukemia virus (BLV) in blood samples quickly, and its sensitivity is 1 fg DNA template, which is 100 times higher than that of PCR. The quick detecting method of bovine leukemia virus provides a quick and accurate diagnostic tool for the clinical diagnosis of bovine leukemia, and has important practical significance for the epidemiological investigation and purification of bovine leukemia.

Compositions and methods are provided for treating certain plant pathogens using microbe-based products. In particular, the subject invention relates to treatment of plant pathogenic viruses, including mosaic virus, as well as plant pathogenic bacteria, using beneficial microbes and/or their growth by-products. In certain embodiments, the growth by-products are biosurfactants.

The invention relates to an antigenic epitope of an African swine fever virus p49 protein and application thereof, and belongs to the field of genetic engineering, the amino acid sequence of the capsid protein of the African swine fever virus p49 (pB438L) protein is used as a material, the antigenic epitope is predicted through an antigenic epitope database, and a corresponding short peptide is synthesized according to the amino acid sequence. African swine fever virus is utilized to infect swine positive serum to screen synthetic peptide capable of performing specific reaction with virus positive serum, antigen epitopes and GST are fused to express recombinant protein, and phages display multi-epitope antigens (virus-like particle display epitopes and VLP) to determine dominant antigen epitopes. Through prediction, immunological screening and identification, the fact that the African swine fever virus p49 protein has 13 epitopes is determined for the first time. The acquisition of the antigen epitopes lays a foundation for research and development of African swine fever detection reagents and kits thereof, preparation of monoclonal antibodies and safe and safe products for identifying and diagnosing strategic prevention and control such as epitope vaccines by taking the antigen epitopes as materials. Not only can huge economic benefits be generated, but also important social benefits are achieved.

The invention discloses a method for quantitatively detecting human T-cell lymphotropic virus provirus, and belongs to the technical field of medical examination. A plasmid for quantitatively detecting the human T-cell lymphotropic virus provirus uses escherichia coli as a vector, and the preservation number of a cloned strain transformed with the plasmid is CGMCC No.19437. The method has the following advantages: the reliable HTLV-1 fluorescence quantitative qPCR method is established, the method is simple to operate, lower in cost, good in specificity, and high in repeatability, the precision, accuracy and linear range can meet clinical testing needs, the sensitivity is high, the minimum detection limit for HTLV-1 is 3 copies/[mu]L, and the method can relatively quantify the pre-viral load of HTLV-1 infected persons, and provide reliable experimental data for prognosis of the infected persons.