29 results about "Replication Process" patented technology

The invention belongs to the technical field of biological medicine, and discloses a nano antibody for resisting porcine delta coronavirus nucleocapsid protein as well as a recombinant, preparation and application of the nano antibody. According to the nano antibody for resisting the porcine delta coronavirus nucleocapsid protein and the recombinant nano antibody thereof, the specific nano antibody 62Nb targeting the PDCoV N protein is obtained through screening, efficient intracellular delivery of the nano antibody 62Nb is realized through engineering modification of a fusion cell-penetrating peptide TAT, and meanwhile, a bacillus subtilis secretory expression system is established to realize large-scale preparation of 62Nb and a recombinant thereof; finally, the replication process of the PDCoV is inhibited in an intracellular expression or exogenous delivery mode. Meanwhile, the invention provides a large-scale preparation method and application of the nano antibody, and provides a new technical scheme for antiviral prevention and detection of PDCoV.

The application provides an application of isotretinoin in preparation of an anti-African swine fever virus drug, and belongs to the technical field of veterinary medicine. It is found that isotretinoin can effectively interfere with the replication process of ASFV by inhibiting the binding of F1055L protein to virus DNA. Experimental data show that isotretinoin can effectively inhibit the replication of ASFV while having low toxicity to host cells. The anti-ASFV method provided by the application effectively inhibits the replication of ASFV through isotretinoin, has high clinical application potential, and provides a new drug strategy for the prevention and treatment of African swine fever.

The invention discloses application of N4-acetylcytidine in prevention and treatment of RNA (Ribonucleic Acid) viruses, belongs to the technical field of biological medicines, and particularly discloses application of N4-acetylcytidine in prevention and treatment of RNA viruses, and the N4-acetylcytidine is used for blocking RNA polymerase replication on which virus RNA depends. The RNA virus genome is easy to mutate, so that the vaccine failure and the off-target of the antiviral drug can be caused, but the action mode of the virus RdRp is extremely conservative, so that the replication process of the virus is inhibited from the RdRp, a new thought is provided for the development of specific and efficient anti-RNA virus drugs, and the application of N4-acetylcytidine is also widened.

The invention belongs to the technical field of biological medicines, and particularly relates to a small molecule compound, a preparation method thereof and application of the small molecule compound in preparation of anti-coronavirus medicines. The small molecule compound provided by the invention shows broad-spectrum anti-coronavirus activity, can effectively inhibit replication, proliferation and virus protein expression of MHV in cells, and resists fatal MHV infection. The compound can also block the duplication process of PEDV in cells, and shows a remarkable anti-PEDV effect. Due to the fact that the MHV and the SARS-CoV-2 are similar in virus structure and transmission mechanism, the antiviral research results aiming at the MHV and the PEDV can provide important reference for developing effective drugs aiming at the SARS-CoV-2 and other coronaviruses. The compounds have important application prospects in the field of development of drugs for resisting coronaviruses such as MHV, PEDV and SARS-CoV-2.

The present invention belongs to the field of molecular biology and veterinary medicine, and provides the use of HSP90 and its inhibitor in regulating the replication of Seneca virus. HSP90 inhibitor can be used to prepare a drug for inhibiting Seneca virus, and HSP90 and its encoding gene can be used to screen a drug for treating or preventing Seneca virus. HSP90 and its inhibitor provided by the present invention can regulate the replication process of Seneca virus, which is of great significance to the pathogenic mechanism and vaccine development of Seneca virus.

The invention discloses lncRNA (long non-coding ribonucleic acid) as well as a coding polypeptide and application thereof. Experiments prove that virus replication can be remarkably inhibited by silencing the lncRNA disclosed by the invention; through overexpression of the lncRNA in cells, virus replication can be significantly promoted after infection of different subtypes of influenza viruses. The lncRNA and the polypeptide GVRP1-ORF thereof play an important role in an influenza virus replication process, provide a new thought and strategy for screening drugs for treating and / or preventing various influenza viruses, provide a new target for treating influenza virus infection, and have a wide application prospect.

The invention discloses an HIV (human immunodeficiency virus) gene regulatory network construction method based on a Boolean network model, which comprises the following steps of: 1, screening HIV-related genes and proteins and regulatory action relationships between genes, between genes and proteins and between proteins from Pubmed, constructing a directed network graph according to the relationships, and optimizing the network; a Boolean polynomial is then defined for each node, and the network is converted into a Boolean model. The invention provides an effective method for researching dynamic information of a network, which is beneficial to revealing a regulation mechanism of a biological system, so that a theoretical basis is provided for disease diagnosis, treatment and drug development. In the aspect of researching a new HIV drug target, the invention also helps to reveal a regulation relation in an HIV infection stage and a replication process, identifies key genes and proteins participating in HIV infection and replication, and helps to discover a new treatment target and optimize an antiviral treatment strategy.

The invention provides application of isotretinoin in preparation of an anti-African swine fever virus medicine, and belongs to the technical field of veterinary medicine. The invention discovers that the isotretinoin can effectively interfere with the replication process of ASFV by inhibiting the combination of F1055L protein and virus DNA (Deoxyribose Nucleic Acid). Experimental data show that the isotretinoin has low toxicity to host cells while effectively inhibiting ASFV replication. According to the anti-ASFV method provided by the invention, the replication of ASFV is effectively inhibited through the isotretinoin, so that the method has relatively high clinical application potential, and a new drug strategy is provided for preventing and treating African swine fever.

Provided herein are analogs of unnatural nucleotides bearing predominantly hydrophobic nucleobase analogs that form unnatural base pairs during DNA polymerase-mediated replication of DNA or RNA polymerase-mediated transcription of RNA. In this manner, the unnatural nucleobases can be introduced in a site-specific way into oligonucleotides (single or double stranded DNA or RNA), where they can provide for site-specific cleavage, or can provide a reactive linker than can undergo functionalization with a cargo-bearing reagent by means of reaction with a primary amino group or by means of click chemistry with an alkyne group of the unnatural nucleobase linker.

The present invention relates to a bacterial continuous evolution system, an orthogonal DNA polymerase and a continuous evolution method. The present invention combines an orthogonal DNA replication system with an orthogonal DNA polymerase to obtain a continuous evolution method that can contain all mutation types, can achieve mutations in long DNA fragments, has good continuity and is easy to operate. By inducing the opening and closing of DNA polymerase expression, the switching between the linear plasmid error-prone mutation process and the high-fidelity replication process is achieved, thereby achieving efficient continuous evolution of the target DNA sequence.

This disclosure concerns compositions and methods for replicating DNA containing a T7 origin of replication (T7 or / ). Compositions and methods herein use a set of proteins that includes a DNA polymerase (DNAP), an RNA polymerase (RNAP), a single-strand binding protein (SSBP) and a helicase-primase (HP), along with one or more linearising proteins and linearising protein-binding nucleic acid sequences to maintain the replicated DNA in a linear conformation. In one example, the addition of a protelomerase recognition sequence (TeIRL) as the linearising protein-binding nucleic acid sequence to a T7 or / plasmid, in combination with the use of a protelomerase (TelN) as the linearising protein during DNA replication, enabled linearisation, replication and maintenance of a T7 or / plasmid in E. coli. The DNAP may be a mutagenic polymerase, so that methods herein may also be used for DNA mutagenesis and evolution of target DNA sequences.

The invention provides a liquid medicine for preventing and treating chicken virus infection, and belongs to the technical field of animal medicine. According to the liquid medicine, raja porosa cartilage polypeptide serves as an active ingredient, the amino acid sequence of the raja porosa cartilage polypeptide is Gly-Glu-Glu-Gly-Thr-Met-Gly-Leu, and a solution with the concentration ranging from 6.25 mu g / mL to 100 mu g / mL is prepared with a PBS buffer solution as a solvent. Experimental results show that the medicine can significantly reduce the cell death rate and chick embryo death rate caused by virus infection and prolong the survival time of infected individuals by preventing H9N2 virus infection and inhibiting the replication process of viruses in host cells. The liquid medicine provided by the invention has the characteristics of high efficiency, safety and easiness in use, and is suitable for virus infection prevention and control of chicken flocks in large-scale farms.

A system and method for performing replication using mediums. A snapshot ‘S’ is selected for replication, and the anchor medium ‘M’ of S is identified. The replica storage array ‘R’ to which M is being replicated determines a list of medium extents which are available. R sends this list to an original storage array ‘O’ and O generates a list of medium extents which need to be sent to R. Only those medium extents corresponding to M which R does not already have will be sent from O to R. Also, R can obtain medium extents from other source storage arrays during the replication process.

The invention belongs to the field of marine gene biological medicine, and particularly relates to siRNA of host metabolic protein and targeted protein interacting with main capsid protein of decapod iridovirus 1 in litopenaeus vannamei and application of the siRNA in inhibition of virus replication. According to the invention, through yeast two-hybrid screening, the host protein which is subjected to specific interaction with the main capsid protein of DIV1 is identified and obtained from litopenaeus vannamei, and the interaction relationship is verified through an in-vitro interaction experiment. Further RNA interference experiment results show that siRNA targeting the host protein can stably play a role in gene knock-down in litopenaeus vannamei bodies, and the knock-down effect can be maintained under the DIV1 infection condition. On the basis, the detection result of the virus load prompts that the replication process of the DIV1 in the litopenaeus vannamei body can be influenced by regulating and controlling the expression level of the host protein.

The present invention discloses the application of plant 14-3-3h protein and its coding gene in resisting Potato virus Y, belonging to the field of biotechnology. The present invention for the first time studies and proves that plant 14-3-3h protein can interact with 6K1 protein participating in the replication process in Potato virus Y; silencing the 14-3-3h gene in plants will promote the infection of PVY; while overexpressing the 14-3-3h gene will inhibit the infection of PVY. Therefore, plant 14-3-3h protein can be used as a new target for resisting Potato virus Y, providing a new idea for the prevention and control of Potato virus Y disease.

The present invention discloses a lncRNA, its encoded polypeptide, and its application. Experiments have confirmed that silencing the lncRNA of the present invention can significantly inhibit viral replication; overexpressing the lncRNA of the present invention in cells can significantly promote viral replication after infection with different subtypes of influenza viruses. The lncRNA of the present invention and its polypeptide GVRP1-ORF play an important role in the influenza virus replication process, providing new ideas and strategies for screening drugs for treating and / or preventing various influenza viruses, and providing a new target for the treatment of influenza virus infection, with broad application prospects.

The present invention belongs to the field of molecular biology and veterinary medicine, and provides the use of HSP70 and its inhibitor in regulating the replication of Seneca virus. HSP70 inhibitor can be used to prepare a drug for inhibiting Seneca virus, and HSP70 and its encoding gene can be used to screen a drug for treating or preventing Seneca virus. HSP70 and its inhibitor provided by the present invention can regulate the replication process of Seneca virus, which is of great significance to the pathogenic mechanism and vaccine development of Seneca virus.

The invention relates to an effect of a VCP inhibitor in preparation of a medicine for treating or inhibiting Ebola virus infection, and belongs to the technical field of biological medicines. Aiming at the Ebola virus, a VCP specific inhibitor NMS-873 is used for remarkably inhibiting the replication and proliferation of the virus, and meanwhile, the expression of the virus protein and the formation process of an inclusion body in the replication process are inhibited. Therefore, the NMS-873 can achieve the purpose of inhibiting Ebola virus infection by remarkably inhibiting Ebola virus infection and release. According to the invention, a new application of the NMS-873 is developed, and a new way for treating or inhibiting the Ebola virus is found.

This disclosure concerns compositions and methods for replicating DNA containing a T7 origin of replication (T7 or / ). Compositions and methods herein use a set of proteins that includes a DNA polymerase (DNAP), an RNA polymerase (RNAP), a single-strand binding protein (SSBP) and a helicase-primase (HP), along with one or more linearising proteins and linearising protein-binding nucleic acid sequences to maintain the replicated DNA in a linear conformation. In one example, the addition of a protelomerase recognition sequence (TeIRL) as the linearising protein-binding nucleic acid sequence to a T7 or / plasmid, in combination with the use of a protelomerase (TelN) as the linearising protein during DNA replication, enabled linearisation, replication and maintenance of a T7 or / plasmid in E. coli. The DNAP may be a mutagenic polymerase, so that methods herein may also be used for DNA mutagenesis and evolution of target DNA sequences.

The present invention belongs to the field of biochips, and in particular relates to a method for converting a gene chip into a printing plate and then mass-producing gene chips. Specifically, taking the finished initial gene chip as the master plate, using DNA polymerase to convert the probe sequences on the master plate into template sequences at corresponding positions on the printing plate according to the principle of base complementary pairing. The probe sequences on the master plate correspond one by one to the template sequences on the printing plate. Then, the template sequences on the printing plate are converted into probe sequences on the new gene chip by using DNA polymerase. The probe sequences on the new gene chip correspond one by one to the template sequences on the printing plate. Through two high-fidelity DNA replication processes catalyzed by DNA polymerase, the probe sequences on the initial gene chip are converted into probe sequences on the new gene chip. The present invention can, like plate-making printing, use the initial gene chip to manufacture a printing plate, and then use the printing plate to mass-produce new gene chips quickly and at low cost.

The invention relates to the technical field of biological detection, in particular to a DNA skin flap controlled FCT-CRISPR detection system and application. The FCT-CRISPR detection system comprises a dumbbell probe containing a DNA skin flap structure, a CRISPR / Cas12a protein, crRNA, a reporter probe and a Cas12a splitting activator SA1, and the dumbbell probe is a dumbbell probe containing a DNA skin flap structure. The DNA skin flap structure comprises a 5 '-end skin flap and a 3'-end skin flap, the 5 '-end skin flap is a Cas12a split activator SA2, and the 5'-end skin flap can be recognized and cut by skin flap endonuclease 1. The detection reaction process adopting the FCT-CRISPR detection system does not need to depend on an additional DNA connection or replication process, and sensitive, accurate and mixed read-and-read detection of the FEN1 activity can be realized.

The present invention relates to the field of biomedicine, and discloses the use of substances that inhibit lnc-ALOX12 in the preparation of influenza virus inhibitors. The present invention for the first time provides that a long non-coding RNA lnc-ALOX12 participates in the replication process of influenza virus as a positive regulatory factor. IAV infection can specifically promote the expression of lnc-ALOX12. Silencing of lnc-ALOX12 can significantly inhibit the transcriptional replication of viral RNA and regulate the nuclear entry of influenza virus PB2 protein. lnc-ALOX12 is involved in host adaptation mediated by PB2 mutants. The present invention reveals that lnc-ALOX12 plays an important role in the replication process of influenza virus and its cross-species transmission, providing a new basis for the development of host lncRNA as an antiviral drug target.

The application belongs to the field of marine gene biological medicine, and particularly relates to a host metabolic protein interacting with a major capsid protein of Decapoda iridovirus 1 in Litopenaeus vannamei, siRNA of the target protein and application of the siRNA in inhibiting virus replication. The application identifies and obtains the host protein specifically interacting with the major capsid protein of DIV1 from Litopenaeus vannamei through yeast two-hybrid screening, and verifies the interaction relationship through in-vitro interaction experiment. Further RNA interference experiment results show that the siRNA targeting the host protein can stably play a gene knockdown role in the body of Litopenaeus vannamei, and maintain the knockdown effect under the condition of DIV1 infection. On this basis, the detection results of virus load suggest that by regulating the expression level of the host protein, the replication process of DIV1 in the body of Litopenaeus vannamei can be affected.

The application provides a use of remdesivir in preparation of a preparation for preventing and / or treating peste des petits ruminants virus, and belongs to the technical field of biological medicine. The application finds that the use of remdesivir in a safe concentration range can significantly inhibit the replication of the peste des petits ruminants virus in host cells, obviously reduce the expression level of virus-related genes and proteins, and exhibit good antiviral activity. Meanwhile, the inhibitory effect of remdesivir on the PPRV presents obvious dose dependence, and with the increase of the acting concentration, the antiviral effect is significantly enhanced, which indicates that the effect is stable and has strong controllability. Moreover, remdesivir mainly acts on the virus replication stage, can effectively block the virus replication process, thereby reducing the virus infection level and reducing the generation of daughter virus particles. The application provides a new technical approach for the prevention and treatment of peste des petits ruminants, makes up for the deficiency of lack of effective antiviral drugs in the prior art, and has a good application prospect.

Gene amplifications are an oncogenic driver utilized by many forms of cancer in tumor development or treatment relapse. Promoting genomic instability or the proclivity to propagate genomic alterations through acquired defects in DNA repair machinery, replication licensing, or cell cycle control, gene amplifications not only drive oncogenesis, but also afford an opportunity for therapeutic exploitation. Here, CRISPR-Cas9 nickases are disclosed which selectively promote cancer cell death in a gene amplification-dependent manner. For example, CRISPR- Cas9 nickases generate a lethal number of highly toxic single-ended double-strand breaks within the genome of proliferating cancer cells harboring amplified genomic loci during DNA replication. Cas9 nickases may serve as a tumor-selective therapeutic that mitigates the collateral damage observed with conventional chemoradiotherapies and avoids chemoresistance.

The application discloses a primer set and a probe for detecting a novel coronavirus 7a subgene, and application and products thereof, and relates to the technical field of biology. The primer set comprises an upstream primer shown as SEQ ID NO. 4 and a downstream primer shown as SEQ ID NO. 2; and the nucleotide sequence of the probe is shown as SEQ ID NO. 3. According to transcriptome sequencing analysis, it is found that the expression amounts of N, E and 7a subgenes are relatively high in the virus replication process. The application provides the primer set and the probe for the novel coronavirus 7a subgene, and realizes rapid detection of the novel coronavirus subgene by using a fluorescent RT-RAA technology, so as to provide a reference for judging the infectivity of a SARS-CoV-2 clinical sample.

The invention belongs to the technical field of biological medicine, discloses an oncolytic virus M1 synergistic composition and application thereof, and particularly discloses application of low-molecular heparin or pharmaceutically acceptable salt thereof in preparation of a product for enhancing the anti-tumor effect of oncolytic virus. The invention provides a technical scheme for improving the treatment effect of the oncolytic virus M1 in tumor treatment through the low-molecular heparin for the first time, the low-molecular heparin can influence the replication process of the oncolytic virus in cancer cells, the combined application of the low-molecular heparin and the oncolytic virus shows a positive effect, a new direction for optimizing the oncolytic virus therapy is opened up, and the application prospect is broad. And a new possibility is provided for improving the treatment effect.

A method of manufacturing a master for use in a wafer-scale replication process is disclosed. The method comprises at least one step of forming a layer of photoresist on a substrate and exposing the layer of photoresist to a radiation pattern to form at least one patterned layer. The method also comprises a step of developing the at least one patterned layer to provide one or more structures defining the master. In an embodiment, the at least one step of forming the layer of photoresist comprises a process of dry film lamination.

The invention uses (i) proximity sequencing (Pro-Seq), in which a fragment is copied redundantly in a manner such that an error introduced during copying does not appear in all copies, and / or (ii) linked target capture (LTC), whereby a nucleic acid is amplified by primers that are linked to target-specific probes to discover and / or detect phased variants as biomarkers.