28 results about "Unstructured Proteins" patented technology

This invention provides a dengue / Zika virus genetically engineered vaccine and its application. The dengue / Zika virus vaccine comprises an open reading frame encoding envelope protein domain III (EDIII) and the non-structural protein NS1, and displays EDIII monomers in the delivery vector shell. Immunization with this vaccine primarily induces a type-specific antibody response, reducing the production of cross-antibodies and thus effectively avoiding or eliminating the risk of antibody-dependent enhancement of infection (ADE). This vaccine can be used to prevent dengue virus and Zika virus infection.

This invention belongs to the field of genetic engineering. By introducing a TC tag between amino acids 199 and 200 of the wild-type bluetongue virus non-structural protein NS2, a recombinant virus was rescued. Analysis of viral plaques and growth curves revealed that the recombinant virus formed plaques of similar size to the wild-type virus, and their growth curves showed no significant difference. Immunofluorescence and FlasH-EDT2 staining of NS2 showed that the fluorescence of NS2 labeled with anti-NS2 antibody overlapped with that labeled with FlasH-EDT2. Three-dimensional imaging of FlasH-EDT2-labeled NS2 revealed that the viral inclusion bodies exhibited fluorescence around the periphery but no fluorescence inside, displaying an overall "core-shell" hierarchical structure. This recombinant bluetongue virus can be used for tracing and localizing NS2 protein and viral inclusion bodies in living cells, and can also be applied to BTV neutralization assays, antiviral drug screening, and other related research.

The invention provides a norovirus genome splicing method and device, a medium and a product. The method comprises the following steps: acquiring a norovirus gene database, wherein the norovirus gene database comprises a non-structural protein coding region sub-library and a capsid protein coding region sub-library; obtaining original sequencing data; performing quality control on the original sequencing data to obtain screened sequencing data; comparing and assembling the screened sequencing data with the non-structural protein coding region sub-library, and determining at least one first recombinant coding region; and comparing and assembling the screened sequencing data with the capsid protein coding region sub-library, and determining at least one second recombinant coding region, and splicing the at least one first recombinant coding region and the at least one second recombinant coding region to obtain a recombinant genome sequence. Through the technical scheme disclosed by the invention, efficient and accurate splicing of the recombinant norovirus genome sequence is realized, and the assembly accuracy and efficiency of the recombinant norovirus genome sequence are improved.

The application discloses a recombinant FCV antigen and a construction method and application thereof. The construction method of the recombinant FCV antigen comprises the following steps: fusing a T cell epitope coding sequence of a non-structural protein NS7 of FCV and a coding sequence of a SpyTag peptide segment through a coding sequence of a linker, then cloning into a baculovirus transfer vector to obtain a recombinant plasmid, and finally integrating the T cell epitope coding sequence into Bacmid through Tn7 transposition to finally obtain the recombinant FCV antigen. The application selects NS7 as a core immunogen, guides the immune system to produce a high cellular immune response, and thus makes up for the deficiency of an existing vaccine in clearing intracellular viruses; meanwhile, a specific T cell epitope is selected in the sequence of NS7 as an immunogen, which can avoid the immunological escape caused by the variation degree of antigens among different strains and virus antigen drift, and thus provides broader protection.

The present application relates to a replicable RNA molecule and the use thereof. The replicable RNA molecule comprises, from the 5′ end to the 3′ end, a 5′ cap, a 5′ UTR, an open reading frame encoding an RNA replicase, a promoter, a sequence of interest, a 3′ UTR and a poly(A) tail, wherein the RNA replicase is capable of amplifying the replicable RNA molecule and is capable of amplifying an RNA molecule containing the sequence of interest and 3′ UTR, wherein the RNA replicase is a nonstructural protein or a functional variant thereof derived from Mosso das Pedras virus (MDPV), Everglades virus (EVEV), Rio Negro virus (RNV), Mucambo virus (MUCV), Highlands J virus (HJV), Pixuna virus (PIXV), Trocara virus (TROV), Cabassou virus (CABV), Tonate virus (TONV), Bebaru virus (BEBV), Fort Morgan virus (FMV), Getah virus (GETV) or Ndumu virus (NDUV).

Synthetic peptides mimicking the nsp10 sequence in the region interacting with nsp16 capable of penetrating cell membranes and inhibiting SARS-CoV-2 replication for the treatment of moderate to severe COVID-19. The invention relates to peptides inhibiting SARS-CoV-2 replication, likely through inhibition of Methyltransferase complexes (NSP10 / NSP16 and NSP10 / NSP14). The peptide of the present invention, P3, contains sequences corresponding to amino acids 89-96 of the non-structural protein 10 (NSP10) of SARS-CoV-2, with the only Cysteine modified to a Methionine. This peptide was made based on two previous designs P1 and P2, which constituted the amino acids 68-96 of the NSP10 protein of SARS-CoV-2.

The invention discloses a protein degradation product and application of non-structural protein NSs of rift valley fever virus or a coding gene of the non-structural protein NSs in preparation of the protein degradation product, and relates to the technical field of protein degradation. On the basis of the property that NSs and recombinant protein of the NSs can autonomously form a fiber-shaped high polymer structure, a degradation machine capable of degrading target protein is self-assembled, so that the specific and efficient degradation of the target protein is realized. Therefore, the new application of the non-structural protein NSs of the rift valley fever virus in targeted degradation of the protein is found, specific efficient degradation of the disease-related protein can be achieved, and the effect of treating diseases is expected to be achieved.

This invention discloses a method for preparing pet stem cell exosomes, belonging to the field of animal medicine technology. This invention treats stem cells with an inducing agent to secrete exosomes with high TFR expression on their surface. These exosomes specifically target the conserved region of the NS1 gene, a non-structural protein of the parvovirus family, allowing the internally loaded siRNA and trefoil factor 3 to target and enter infected cells, directly inhibiting viral replication at the gene level to treat feline panleukopenia virus (FPV) and canine parvovirus infections and alleviate sequelae. This invention solves the problems of existing technologies, such as difficulty in targeting, lack of specific drugs, reliance on intensive supportive care leading to a short treatment window, inability to directly clear the virus, risk of cross-infection, and potential long-term sequelae. It has the potential to be applied to the preparation of drugs for treating canine parvovirus and feline panleukopenia.

The invention discloses a recombinant FCV antigen and a construction method and application thereof. The construction method of the recombinant FCV antigen comprises the following steps: fusing a T cell epitope coding sequence derived from a non-structural protein NS7 of FCV with a coding sequence of a SpyTag peptide fragment through a coding sequence of a linker, and cloning to a baculovirus transfer vector to obtain a recombinant plasmid; and finally, integrating the T cell epitope coding sequence into Bacmid through Tn7 transposition to finally obtain the recombinant FCV antigen. According to the invention, NS7 is selected as a core immunogen to guide an immune system to generate high cellular immune response, so that the defect of an existing vaccine in removing viruses in cells is overcome; meanwhile, T cell epitopes with specificity are selected from the sequence of NS7 as immunogens, so that the variability of antigens among different strains and immune escape caused by virus antigen drift can be avoided, and wider-spectrum protection is provided.

The invention discloses a rotavirus reassortment attenuated strain LG12 and application of the rotavirus reassortment attenuated strain LG12. The rotavirus reassortment attenuated strain LG12 comprises: i) a coding gene of a structural protein VP7 of a human G12 type rotavirus strain G12P [6]; and ii) coding genes of structural proteins VP1, VP2, VP3, VP4 and VP6 and non-structural proteins NSP1, NSP2, NSP3, NSP4 and NSP5 of the sheep rotavirus strain LLR. The reassortment strain provided by the invention has the safety of animal strains and the immunogenicity of human G12 type RV, can effectively prevent infection of human G12 type wild strains, and can have different degrees of cross protection on infection of other various G type RV wild strains; the method has great significance for developing RV vaccines which cover more G and P genotypes of Chinese epidemic RVA, have stronger prevention pertinence and provide a better protection effect on Chinese infantile diarrhea.

The application discloses an epitope tandem polypeptide of porcine reproductive and respiratory syndrome and related biological materials and application thereof, and belongs to the field of viral antigen vaccines or biomedical technologies. The application provides an epitope tandem polypeptide which is composed of neutralizing antigen epitopes, Th epitopes and CTL epitopes of multiple structural proteins and non-structural proteins of PRRSV in series, and includes PNB, PTH and PCTL. The epitope tandem polypeptide provided by the application can be directly used for preparing a subunit vaccine of PRRSV, and has the advantages of simple preparation method, strong immunogenicity and good safety. The epitope tandem polypeptide can also be combined with an inactivated vaccine of PRRSV and applied to enhance cellular immunity induced by the inactivated vaccine, so that the epitope tandem polypeptide can more effectively prevent and protect pigs from PRRSV infection. The epitope tandem polypeptide composition contains common epitope sequences of seven representative strains (LV, VR-2332, CH-1a, JXA1, NADC30, NADC34 and RFLP 1-4-4) of PRRSV, and is expected to have good cross-protection for infection of different strains.

The application discloses a recombinant novel goose parvovirus infectious clone plasmid and application, the infectious clone plasmid includes the full-length genome of novel goose parvovirus, and a synonymous mutation marker is introduced at the cysteine codon position in the non-structural protein coding region, under the condition that the amino acid sequence is not changed and the offspring virus replication and infection ability are not influenced, a stable single nucleotide genetic marker is provided for the novel goose parvovirus.The infectious clone plasmid can be used for rescuing the recombinant virus carrying a specific synonymous mutation marker, the obtained recombinant virus has good duck embryo adaptability and cell infection ability, and is suitable for application scenes such as cell infection and immunofluorescence detection.The application provides an important technical tool for novel goose parvovirus genetic marker tracking, pathogenic mechanism research, genetic variation analysis and vaccine research and development.

The identified peptide sequences form moderate to strong heterodimeric complexes with SARS-COV2 (B.1.617.2) virus Spike envelope proteins and a range of virus nonstructural protein sequences NSP1-14. These identified peptides are proposed to use in antiviral immunotherapy and supportive combination with chemical compound(s) for chemotherapy. The sequence ID of the identified peptides are, Seq1: ISD1, Seq2: ISD2, Seq3: ISD3, Seq4: ISD4, Seq5: ISD5, Seq6: ISD6, Seq7 Ip2, Seq8: Ip1, Seq9: Ip3, Seq10: Ip5, Seq11: Ip7, Seq12: Ip8, Seq13: Ip9, Seq14: SOI1, Seq15: SOI2, Seq16: SOI5, Seq17: SOI7, Seq18: SOI9, Seq19: SO10, Seq20: SO11.

The invention belongs to the technical field of virus construction, and particularly relates to a bovine parainfluenza virus type 3 V protein deletion strain and application thereof. The construction process of the strain comprises the following steps: by taking pBPIV3-SX full-length infectious clone plasmid as a template, respectively amplifying two gene segments before and after a base editing site of a V gene, and fusing the obtained amplified segments to obtain a segment containing V gene deletion; and replacing a fragment containing V gene deletion to BssHII and PacI restriction enzyme cutting sites of the pBPIV3-SX plasmid, and transfecting to carry out virus rescue, thereby obtaining the recombinant plasmid pBPIV3-SX. According to the invention, on the basis of a reverse genetics operation technology, a V protein deleted infectious clone plasmid is constructed, and a recombinant deleted virus is successfully rescued; meanwhile, the V protein is obtained through an escherichia coli expression system, a mouse is immunized to obtain a polyclonal antibody, a theoretical basis is provided for functional research of the BPIV3 non-structural V protein and development of genetic engineering vaccines, and the BPIV3 non-structural V protein has important scientific value and application value.

The invention relates to the technical field of antibodies, in particular to a 3D protein monoclonal antibody and application thereof. The monoclonal antibody comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprises the following three complementary determining regions CDR: CDR1 shown as SEQ ID No.5, CDR2 shown as SEQ ID No.6, and CDR3 shown as SEQ ID No.7; the heavy chain variable region comprises the following three complementary determining regions: CDR1 shown as SEQ ID No.5, CDR2 shown as SEQ ID No.6, and CDR3 shown as SEQ ID No.7; the light chain variable region comprises the following three complementary determining regions CDR: CDR1 as shown in SEQ ID No.8, CDR2 with the sequence of WAS, and CDR3 as shown in SEQ ID No.9. The light chain variable region has the advantages that the light chain variable region can be used for preparing the light chain variable region; the invention provides a specific monoclonal antibody of a foot and mouth disease virus non-structural protein 3D protein. A Western blotting test shows that the specific monoclonal antibody specifically reacts with the non-structural protein 3D; indirect immunofluorescence tests show that the antibody has good reactivity with an FMDV O-type strain.

The present disclosure relates to polypeptides that specifically bind to Dengue virus non-structural protein 1, including antibodies and fragments thereof. The antibody or antigen-binding fragment thereof may specifically bind Dengue virus (DENV) serotype 4 and include: a heavy chain variable region that comprises at least one CDR amino acid sequence selected from the group consisting of: SGYNWH, YIHYSGGTNYNPSLKS, RTGTVPFAY, SYVMH, YLNPYNDDTKYNEKFKG, and GPPYALDY. The present disclosure further relates to methods of producing the polypeptides of the present disclosure, methods of diagnosing DENV, and methods of treating a DENV infection.

This invention provides an optimized method for enhancing self-amplifying RNA expression and an enhanced self-amplifying RNA. Specifically, it provides an optimized method for enhancing self-amplifying RNA expression, which involves codon optimization. The optimized region is a specific region of the alphavirus non-structural protein sequence of the self-amplifying RNA, where the alphavirus non-structural protein is nsP1234. It also provides a method for preparing the enhanced self-amplifying RNA, an enhanced self-amplifying RNA, and a plasmid. This application improves the expression efficiency of the self-amplifying RNA target gene by designing and constructing codon sequence optimization for certain regions of these non-structural proteins.

The invention discloses a prawn culture pathogenic bacteria multiple ERA detection primer and probe based on a biochip technology and application of the prawn culture pathogenic bacteria multiple ERA detection primer and probe. The kit is characterized in that the primers and the probe respectively aim at tcdB genes of highly pathogenic vibrios, hypothetical protein genes of white spot syndrome viruses, 16S rRNA genes of enterocytozoon hepatopenaei, non-structural protein genes of infectious subcutaneous and hematopoietic necrosis viruses, DNA primer enzyme genes of decapod iridovirus 1 and PirB genes of bacterial prawn acute hepatopancreatic necrosis pathogens; the method comprises the following steps of: detecting a capsid protein gene of a prawn infectious muscle necrosis virus, a capsid protein gene of a nedavirus, a capsid protein gene of the nedavirus, a structural protein ORF1 gene of the prawn infectious muscle necrosis virus, a non-structural polyprotein gene of a taura syndrome virus and a Rep-1b gene of a yellow head virus, and further discloses specific sequences. The sensitivity, the specificity and the accuracy are high.

The present application relates to a method for extracting epitopes effective in the prevention or treatment of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the method comprising the steps of: (a) extracting, from an anti-spike antibody database, first structural data including known structures and second structural data including unknown structures, and extracting, from a SARS-CoV-2 proteome database, third structural data including known structures; (b) carrying out 3D modeling on the second structural data, and classifying the third structural data into a first group including structural proteins and a second group including both structural proteins and non-structural proteins; (c) carrying out spike-antibody docking on the basis of the first group and the second structural data on which 3D modeling has been carried out; (d) constructing a spike-antibody database on the basis of the first structural data and the docked data, and predicting conformational epitopes on the basis of the second group; (e) characterizing epitopes on the basis of the spike-antibody database and the conformational epitopes; and (f) selecting final epitopes on the basis of analysis results.

The present invention provides live attenuated vaccines for the prophylactic treatment of Chikungunya virus infections. The vaccines comprise a genetically modified Chikungunya virus that has a reduced replication capacity due to a mutation or deletion in the R5 region of the hypervariable domain of nsP3, a nonstructural protein that interacts with host factors. The inventors have indeed discovered that the R5 region of the hypervariable domain of nsP3, a nonstructural protein that interacts with host factors, is critical for CHIKV replication and pathogenesis. Specifically, the inventors have shown that the R5 region is required for the interaction of nsP3 with FHL1, BIN1 and CD2AP, three host proteins that are involved in the regulation of actin cytoskeleton and membrane trafficking. These host proteins are expressed in muscle and joint tissues, which are the main targets of CHIKV infection and inflammation. By mutating or deleting the R5 region of nsP3, the inventors have generated a genetically modified CHIKV (CHIKV-ΔR5) that has a reduced replication capacity and virulence compared to the WT virus. Moreover, the inventors have demonstrated that CHIKV-ΔR5 is able to elicit a strong neutralizing antibody response and protect mice from lethal challenge with WT virus. The present invention also provides screening methods for identifying test substances that are capable of inhibiting the interaction between nsP3 and FHL1, CD2AP and BIN1, wherein the selected test substances would be suitable for the treatment of CHIKV infections.

The present invention provides compositions and methods for making circular RNAs, in particular circular RNAs encoding one or more proteins and having superior stability. Such circular RNAs comprise complementary regions that are capable of forming double-stranded regions that are important for maintaining the stability of the circular RNA under different environments and that facilitate translation and circularization. Such complementary regions can be formed by codon optimization or can be introduced as heterologous sequences. The present invention also provides circular RNAs that express self-amplifying RNA non-structural proteins, e.g., (nsP1-4), which can amplify independent mRNA molecules containing corresponding conserved sequence elements (CSEs). These mRNAs can encode a gene of interest (GOI). When they are introduced together into a target cell, the circular RNAs can amplify the mRNAs, thereby enabling stable high expression of the gene of interest.

Provided herein are live attenuated viruses for protection against respiratory syncytial virus (RSV) and / or coronavirus Sars-CoV-2. The live attenuated chimeric virus strains utilize a master backbone based on a live attenuated influenza virus (LAIV), which includes a deletion of the viral virulence element, the NS1 (non-structural protein 1) (DeLNS1). These chimeric strains are engineered to express one or more antigens of RSV alone or in combination with Sars-CoV-2. The chimeric virus strain can protect a subject in need thereof against a challenge from any of RSV, Sars-CoV-2, influenza, or a combination thereof. This viral vector system offers an important strategy for developing highly attenuated and immunogenic live attenuated vaccines with the capacity to induce protective immunity against the three respiratory infections.

Pharmaceutical compositions of the invention comprise diazepines derivatives having a disease-modifying action in the treatment of diseases associated with biological effect that include disease state, and any disease type / class involving biological effect.

The present invention relates to a recombinant nonstructural protein 1 (NS 1), a recombinant influenza virus and an immunological composition including the same, as well as a method of treating or preventing a disease or condition caused by or associated with an influenza virus in a subject in need thereof. At least one amino acid residue is mutated or deleted in 4 contiguous amino acid residues of TRAF3-interacting motifs (TIMs) within an effector domain of the recombinant NS 1. A recombinant influenza virus including the recombinant NS1 is comparable to the wild-type influenza virus in virus replication ability, and the recombinant influenza virus can elevate interferon activation of a subject for achieving better immune response and better immunological protection, leading in a method of treating or preventing a disease or condition caused by or associated with an influenza virus in a subject in need thereof.

The invention discloses variable region amino acid sequences of two monoclonal antibodies of yellow fever virus non-structural protein NS1 and application of the variable region amino acid sequences, and belongs to the technical field of medicines. Yellow fever virus non-structural protein NS1 expressed by human embryo kidney 293 cells is used as an antigen to immunize a rabbit, B cells capable of being specifically combined with the yellow fever virus non-structural protein NS1 are screened from rabbit spleen cells through flow sorting, and signal peptide and variable region gene fragments of an antibody are cloned through reverse transcription-polymerase chain reaction. The method comprises the following steps of: constructing a monoclonal antibody, connecting the monoclonal antibody and a constant region gene into an expression vector, expressing and purifying by mammalian cells, and detecting by enzyme-linked immunosorbent assay to obtain two monoclonal antibodies with high affinity and independent epitopes, and the two monoclonal antibodies are not combined with non-structural proteins NS1 of other eight viruses of flaviviridae flaviviruses. The two monoclonal antibodies have application value in diagnosis and prevention of yellow fever.

Provided herein is an isolated polynucleotide, which encodes alphavirus non-structural proteins nsp1, nsp2, nsp3 and nsp4 and a polypeptide comprises CD8+ T cell epitope. Provided herein is also a bivalent alphavirus replicon vaccine, which is a combination of a first polynucleotide which encodes alphavirus non-structural proteins nsp1, nsp2, nsp3 and nsp4 and an antigenic peptide and a second polynucleotide which encodes alphavirus non-structural proteins nsp1, nsp2, nsp3 and nsp4 and a CD8+ T cell epitope. The vaccine is useful against virus infection, especially, COVID-19 or SARS-CoV-2 infection, the treatment of a cancer and / or an inflammatory disease.

The application belongs to the field of genetic engineering, and particularly relates to a recombinant blue tongue virus which fuses and expresses a tetracysteine (TC) tag on NS2 protein and a construction method. The application inserts a TC tag at the C terminal (after the 354th amino acid) of the non-structural protein NS2 of a wild type blue tongue virus to construct a recombinant blue tongue virus BTV1-NS2-354TC. The recombinant blue tongue virus has no obvious difference from the wild type blue tongue virus in biological characteristics, and the NS2 protein expressed by the recombinant blue tongue virus and carrying the TC tag can be dyed by a double arsenic dye (FlAsH-EDT2), shows green fluorescence under a fluorescence microscope, and can be used for visual quantitative detection of BTV infection and positioning and tracing research of the NS2 protein.

The invention discloses a recombinant serum type 3 duck adenovirus and a construction method and application thereof, the recombinant serum type 3 duck adenovirus replaces a non-structural protein gene ORF52 in the serum type 3 duck adenovirus with an exogenous target gene or inserts the exogenous target gene into the non-structural protein gene ORF52, and the nucleotide sequence of the non-structural protein gene ORF52 is shown as SEQ ID NO. 1. The recombinant serum type 3 duck adenovirus for expressing green fluorescent protein is constructed by targeting an exogenous gene EGFP to a non-structural protein gene ORF52 of the duck adenovirus type 3 by utilizing a gene editing technology, and the non-structural protein gene ORF52 is proved to be capable of serving as an insertion site of the exogenous gene, so that the recombinant serum type 3 duck adenovirus can be used as an insertion site of the exogenous gene. And a foundation is laid for developing a genetic engineering recombinant DAdV-3 multi-combined multivalent vaccine based on taking DAdV-3 as a virus vector.