122 results about "Viral genomes" patented technology

The invention discloses a method for quickly rescuing bovine coronavirus epidemic strains and application of the method. The bovine coronavirus SHZ isolate is subjected to efficient segmented cloning, the obtained segment and a Linker sequence are subjected to a cyclic polymerase extension reaction, and the obtained reaction product can be directly transfected to 293T cells for virus packaging without purification. Different from a traditional method, the method does not need to amplify full-length virus cDNA clone by means of bacteria and yeast, but directly obtains a sufficient amount of preliminary products through PCR, and avoids the problems of instability and low efficiency when partial sequences of a virus genome proliferate in a bacteria or yeast host. By adopting the reverse genetic system, the recombinant bovine coronavirus expressing the foreign protein is successfully rescued, a brand new technical platform is provided for dynamic visualization research of in-vivo and in-vitro replication of the virus, and an efficient and flexible tool is also provided for virology research, development of virus vector vaccines and screening of antiviral drugs.

A CVB virus expression vector comprising a PIV5 W3A viral genome comprising a mutation at amino acid residue S157 or S156 of the P / V gene and a deletion of the small hydrophobic (SH) gene of the PIV5 W3A viral genome, wherein the amino acid substitution at amino acid residue S157 or S156 comprises a substitution of serine (S) with phenylalanine (F) or asparagine (N), and the SH gene has a deletion of the SH open reading frame or the entire SH gene transcription unit. The CVB virus expression vector expresses a heterologous polypeptide, including SARS-CoV-2 spike (S), and / or nucleocapsid (N) and / or membrane (M) proteins, RSV fusion protein (F), or other antigens.

The invention provides a capture primer group and a kit for targeted Salivirus genome sequencing and application of the capture primer group and the kit, and belongs to the technical field of genome sequencing. The targeted capture primer group of the Salivirus virus genome comprises a primer 1 and a primer 2, the nucleotide sequence of the primer 1 is as shown in SEQ ID NO. 1, and the nucleotide sequence of the primer 2 is as shown in SEQ ID NO. 2. According to the invention, a universal reverse transcription oligo-T primer and the Salivirus specific primer group provided by the invention are put into the reverse transcription step at the same time, so that not only is a macro transcription effect achieved, but also a Salivirus sequence in a sample is synchronously identified for specific capture. According to the method, the complete Salivirus genome sequence can be efficiently obtained, and the method has good coverage and sequencing adaptability to the virus genome.

Nucleotide compositions, recombinant vectors, and recombinant viral genomes are disclosed that include a modified horseradish peroxidase (HRP) gene sequence, as well as recombinant viruses that have the modified HRP gene sequence incorporated into the viral genome thereof. The recombinant viruses are utilized in an HRP-based assay to analyze the neutralization potential of candidate antiviral agents, such as antiviral antibodies.

Provided herein are polynucleotides comprising a coding sequence for expression of a fibroblast growth factor 21 (FGF21) pathway activating agent. Also provided are vectors, recombinant viral genomes, and recombinant virus compositions comprising said polynucleotides, as well as associated methods.

The invention relates to a helper adenovirus genome wherein expression of one or more of the adenoviral early E1 genes, E2 genes including DNA-binding protein (DBP), pre-terminal protein (pTP) and DNA polymerase, or production of functional proteins are regulated. The invention also relates to a process for delivery of the helper-dependent or gutless adenovirus vector genome and modulating replication of the helper virus DNA in the manufacturing cells, in which to allow a sufficient length of time before excision, cutting, or recombination to removal of the viral packaging signal DNA sequence from the helper adenovirus genome prior to induction of viral DNA replication, and expression of helper proteins that are required for adenovirus capsid particle formation and packaging, or encapsidation, of the helper-dependent or gutless adenovirus vector genomes, while preventing the packaging, or encapsidation, of the helper adenoviral genome into preform adenoviral capsids.

This invention discloses a chikungunya virus genome fragment armored RNA reference, its preparation method, and its application. The preparation process involves the following steps: (1) constructing the prokaryotic expression backbone vector pACYCAR; (2) cloning the CKV genome fragment CKA (nt17-2276); (3) constructing the prokaryotic expression vector pACYCAR-CKA; (4) inducing the expression of armored RNA; and (5) purifying the armored RNA. This invention solves the technical problems of high biosafety risk, poor stability, and insufficient gene fragment coverage in existing CKV nucleic acid detection references. It fills the gap in the lack of high-quality references for CKV detection.

The invention discloses a primer group and a kit for detecting a new bunyavirus genome and application of the primer group and the kit. The primer group comprises a primer group I and a primer group II, wherein the primer group I comprises nucleotide sequences as shown in SEQ ID NO.1-39; and the primer group II comprises a nucleotide sequence as shown in SEQ ID NO.40-77. The primer group I comprises a nucleotide sequence as shown in SEQ ID NO.7. The primer group can be used for amplifying the whole genome of various strains of the new bunyavirus. According to the application, nucleic acid of the new bunyavirus in a sample is obtained by using an efficient and stable nucleic acid extraction technology, cDNA is synthesized by using an optimized reverse transcription system, then two multiple PCR primer pools are designed by using an imbricate amplification principle, whole-genome segmented amplification is performed on the cDNA by using high-fidelity enzyme, and the cDNA is amplified by using high-fidelity enzyme to obtain the new bunyavirus. And finishing the detection of the new bunyavirus genome in combination with a three-generation sequencing platform. The method can accurately analyze nucleotide mutation through sequence alignment, has the characteristics of simplicity and convenience in operation, high efficiency, rapidness, high specificity and high accuracy, and provides a powerful tool for monitoring, prevention and control and research of the new bunyavirus.

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 invention discloses a nucleic acid molecule which comprises an open reading frame for coding an EBV antigen, and the EBV antigen is a specific antigen which is contained in a diseased cell of an EBV related disease, is coded by an EBV virus genome and is in a cell and on a cell membrane; the EBV antigen comprises LMP1, LMP2A, LMP2B, EBNA-LP, EBNA1, EBNA2, EBNA3A, EBNA3B, EBNA3C, EBNA4, EBNA5, EBNA6, EBNA7 and EBNA8, as well as lipid nanoparticles containing nucleic acid molecules, a pharmaceutical composition and application thereof, and belongs to the technical field of biology. The nucleic acid molecule, the mRNA, the composition containing the nucleic acid molecule, and the lipid nanoparticles can significantly increase the number of tumor specific immune cells, effectively activate immune response, and significantly inhibit tumor growth, and have significant anti-tumor activity for nasopharynx cancer and lymphoma.

The present disclosure provides materials and methods for detecting the presence of a target, such as a nucleic acid, biomolecule or biological particle, or virus genome in a sample. The present disclosure further provides point of care detection methods that cane done quickly and simply, optionally without sophisticated instrumentation. The present disclosure further provides improved methods for detecting and quantifying detection signals from various emulsions.

The disclosure relates to polynucleotides, e.g., viral genomes, encoding syntaxin-binding protein-1 (STXBP1), vectors, e.g., adeno-associated virus (AAV) particles, comprising said polynucleotides, compositions comprising said polynucleotides or vectors, and methods of making or delivering to a cell or subject. The polynucleotides, vectors, compositions, and methods of the present disclosure are useful for the treatment of subjects who have, have been diagnosed with having, or are at risk of having a STXBP1-related disorder, e.g., STXBP1 Developmental and Epileptic Encephalopathy (DEE) and / or other STXBP1-related disorders, or at least one symptom thereof.

Described herein are recombinant viral genomes comprising a nucleotide sequence encoding a telomerase reverse transcriptase (TERT) operably linked to a tissue-specific and / or organ-specific promoter. Aspects described herein may be used in the treatment of conditions associated with shortened telomere length, such as pulmonary fibrosis, myocardial infarction and conditions associated therewith.

Described are optimized RNAi agents, compositions that include RNAi agents, and methods for inhibition of coronavirus (CoV) viral genome. The optimized CoV RNAi agents and RNAi agent conjugates disclosed herein inhibit the expression of a SARS-CoV-2 viral genome, and the targeted portions of the genome are conserved across a variety of known coronaviruses. Pharmaceutical compositions that include one or more optimized CoV RNAi agents, optionally with one or more additional therapeutics, are also described. Delivery of the described CoV RNAi agents to pulmonary cells, in vivo, provides for inhibition of CoV viral genome expression, including SARS-CoV-2, which can provide a therapeutic benefit to subjects, including human subjects, for the treatment of various diseases including COVID-19.

The present disclosure relates to constructs, in particular lentiviral constructs, for use in expression systems for cellular and / or gene therapy applications. These constructs include viral genomes whose expression is controlled by the RNA polymerase I (Pol I) promoter.

This disclosure concerns methods of analysing and sequencing viral genomic DNA, in particular Epstein-Barr virus (EBV) genomic DNA, using viral-specific nucleic acid capture probes and long read sequencing.

The present invention is directed to a method for detecting the presence or absence of monkeypox virus (MPXV) in a biological sample, the method comprising: a) contacting the sample or nucleic acid extracted from said sample with reagents and an oligonucleotide combination capable of amplifying a target region of a monkeypox viral genome, wherein said oligonucleotide combination is selected from the group consisting of: i) a B7R oligonucleotide primer set specific for amplifying a target sequence in the B7R gene of the MPXV genome generating an MPXV amplification product comprising the sequence of SEQ ID NO:65; and ii) a G2R oligonucleotide primer set specific for amplifying a target sequence in the G2R gene of the MPXV genome generating an MPXV amplification product comprising the sequence of SEQ ID NO:63; b) performing an in vitro nucleic acid amplification reaction, wherein any MPXV nucleic acid present in the sample is used as a template to generate the MPXV amplification product corresponding to the oligonucleotide combination of i) or ii); and c) detecting the presence or absence of the MPXV amplification product generated in step b), wherein the presence of the MPXV amplification product is indicative of the presence of MPXV in the biological sample. The present invention also provides oligonucleotides which can be used in the method of the invention as primers and probes.

The application discloses a kind of establishment and application of flavivirus replication-dependent RNA recombination experimental system, belong to the field of biotechnology.The RNA recombination system includes two components: tanbusu virus E gene deletion subgenomic replicon CQW1-ΔE, and the defective virus genome CQW1-ΔNS1 or MM1775-ΔNS1 missing partial NS1 gene sequence.The two components cannot produce progeny virus alone, but after co-transfection, due to RNA recombination, a large number of recombinant progeny viruses can be produced within a few days after transfection, the recombination efficiency is extremely high, and continuous passage is not required, greatly shortening the construction cycle and saving production costs;At the same time, the progeny virus can form plaques on cells, and the efficiency of RNA recombination can be quantitatively measured by plaque counting, and the key factors affecting tanbusu virus RNA recombination can be analyzed.

The invention relates to a helper adenovirus genome wherein expression of one or more of the adenoviral early E1 genes, E2 genes including DNA-binding protein (DBP), pre-terminal protein (pTP) and DNA polymerase, or production of functional proteins are regulated. The invention also relates to a process for delivery of the helper-dependent or gutless adenovirus vector genome and modulating replication of the helper virus DNA in the manufacturing cells, in which to allow a sufficient length of time before excision, cutting, or recombination to removal of the viral packaging signal DNA sequence from the helper adenovirus genome prior to induction of viral DNA replication, and expression of helper proteins that are required for adenovirus capsid particle formation and packaging, or encapsidation, of the helper-dependent or gutless adenovirus vector genomes, while preventing the packaging, or encapsidation, of the helper adenoviral genome into preform adenoviral capsids.

The invention discloses a coronavirus attenuated strain, a vaccine as well as a preparation method and application of the coronavirus attenuated strain. Compared with a natural strain, the coronavirus attenuated strain disclosed by the invention at least comprises deletion of a No.5 stem loop (Stem-loop 5, SL5) from the 131 site to the 350 site of a 5 '-non-coding region (5' UTR) or mutation of any nucleotide maintaining a stem loop structure in a second stem loop from the 175 site to the 299 site of the SL5; compared with a natural strain, the virulence of the SL5-deleted mutant virus strain is obviously reduced; the SL5-deficient mutant virus strain has better genetic stability, and the carried virus genome has no risk of being integrated into the host cell genome. Therefore, the coronavirus attenuated strain disclosed by the invention can be used as an attenuated live vaccine for preventing coronavirus infection, and has a good application prospect.

The invention relates to the technical field of medicine and biology, and particularly discloses a stress-inducible virus and application thereof.The stress-inducible virus comprises a stress-inducible adenovirus or a stress-inducible lentivirus, the nucleotide sequence of the stress-inducible adenovirus or the stress-inducible lentivirus is shown as SEQ ID NO: 1, and the nucleotide sequence of the stress-inducible adenovirus or the stress-inducible lentivirus is shown as SEQ ID NO: 2. The preparation method comprises the following steps: constructing a virus vector, wherein the virus vector has a nucleotide sequence as shown in SEQ ID NO: 1; and virus packaging: transfecting HEK293T cells by adopting shuttle plasmids containing SEQ ID NO: 1 and skeleton plasmids containing viral genome sequences, and collecting a virus stock solution to obtain the stress induced virus. The invention also discloses application of the stress-induced virus in preparation of medicines for preventing and treating oral tumors. Controllable release of the foreign protein under a specific signal is realized. The therapeutic protein produced by the invention can maximize the curative effect of drugs such as CD47 targeted protein and the like, and meanwhile, the toxic and side effects of the drugs on other tissues of the whole body are reduced.

This invention belongs to the fields of microbial technology and veterinary pharmacology, and discloses a strain of *Alternaria alternata* isolated from Artemisia argyi, its fermentation supernatant, and its application in the preparation of anti-PRRSV drugs. The strain's preservation number is CCTCC NO: M2026443. The fermentation supernatant of this strain can inhibit PRRSV infection of cells and inhibit viral proliferation on cells. Cell-level anti-infection studies show that this strain has significant anti-PRRSV activity in vitro, reducing viral titer, viral genome copy number, and viral nucleoprotein production. It can be used to prepare a broad-spectrum anti-porcine reproductive and respiratory syndrome virus drug to control the occurrence of porcine reproductive and respiratory syndrome.

The invention relates to the technical field of virus detection, in particular to a primer probe combination, a kit and a method for detecting G3 gene group pig bocavirus based on a constant-temperature RAA fluorescence method in a non-diagnostic purpose mode. Aiming at a conserved region of a PBoV G3 virus genome, a probe which is specially modified and designed is adopted and has a synergistic effect with an upstream primer as shown in SEQ ID NO.4 and a downstream primer as shown in SEQ ID NO.7. The primer probe combination is used for detecting the G3 gene group pig bocavirus by a constant-temperature RAA fluorescence method, the detection sensitivity and specificity can be remarkably improved, viral nucleic acid as low as 10.5 copies / mu L can be detected in an extremely short time, meanwhile, cross reaction with other pig bocavirus gene groups or common pig pathogens is effectively avoided, and the detection sensitivity and specificity are improved. And a dual detection mode of instrument quantification and blue light naked eye interpretation can be supported, so that the detection efficiency and the convenience of field application are effectively improved.

The present invention provides: an oncolytic virus such as a conditionally replicating adenovirus carrying the CXCL10 gene; the oncolytic virus additionally carrying the IL-2 gene and / or the GM-CSF gene on the same viral genome; a combination of the oncolytic virus and a separate oncolytic virus carrying the IL-2 gene and / or the GM-CSF gene; and a cancer treatment agent comprising any of the aforementioned oncolytic viruses or a combination thereof as an active ingredient.

The disclosure relates to adeno-associated (AAV) particles comprising viral genomes encoding cyclin-dependent kinase-like 5 (CDKL5) proteins and peptides, compositions comprising said AAV particles, and methods for making and delivering said AAV particles to a cell or subject. The AAV particles, compositions, and methods of the present disclosure are useful for the treatment of subjects who have, have been diagnosed with having, or are at risk of having a CDKL5 deficiency disorder (CDD), developmental and epileptic encephalopathy 2, atypical Rett syndrome, and / or other CDKL5-related disorders or at least one symptom thereof.

Provided herein arc polynucleotides comprising a coding sequence for expression of a fibroblast growth factor 21 (FGF21) pathway activating agent. Also provided are vectors, recombinant viral genomes, and recombinant virus compositions comprising said polynucleotides, as well as associated methods.

To provide a method for accurately and quantitatively detecting human norovirus having activity.SOLUTION: A method for detecting a human norovirus according to the present invention is a method for detecting a human norovirus having an activity present in a test object, the method including the steps of preparing a magnetic bead-bound body in which an anti-human norovirus IgY antibody is bound to a surface of a magnetic bead, mixing the test object containing the human norovirus with the magnetic bead-bound body, collecting the magnetic bead-bound body, separating an RNA virus genome of the human norovirus bound to the magnetic bead-bound body, synthesizing cDNA from the separated RNA virus genome and amplifying the cDNA, and determining that the human norovirus having the activity is present when an amount of an amplification product obtained in the amplifying step is higher than a reference value.SELECTED DRAWING: Figure 1

The present disclose provides a primer, probe, and kit for broad-spectrum detection of mycobacteria and use thereof, relating to the technical field of broad-spectrum detection of mycobacteria; the primer-probe combination including forward primer, reverse primer and probe, the nucleotide sequence of forward primer is shown in SEQ ID NOs. 1 and 2, the nucleotide sequence of reverse primer is shown in SEQ ID NO. 3, and the nucleotide sequence of detection probe is shown in SEQ ID NOs. 4-6. The primer-probe combination provided by the present disclose can detect more than 160 types of mycobacteria, covering a wide range; the detection time is greatly shortened; having strong specificity, and has no cross reaction with non-mycobacterial cells and viral genomes; having high detection sensitivity, with the detection limit reaching 100 CFU / ml and the genome detection limit being 10-20 copies / reaction; having high durability and high resistance to matrix interference.

A CVB virus expression vector comprises a PIV5W3A virus genome, the PIV5W3A virus genome contains a mutation at an amino acid residue S157 or S156 in a P / V gene of the PIV5W3A virus genome and a deletion of a small hydrophobic (SH) gene, the amino acid substitution at the amino acid residue S157 or S156 comprises that serine (S) is substituted by phenylalanine (F) or asparagine (N), and the amino acid substitution at the amino acid residue S157 or S156 is substituted by phenylalanine (F) or asparagine (N). And wherein the SH gene deletes an SH open reading frame or deletes the entire SH gene transcription unit. CVB virus expression vectors wherein the vectors express heterologous polypeptides comprising SARS-CoV-2 spike (S) and / or nucleocapsid (N) and / or membrane (M) proteins, RSV fusion proteins (F) or other antigens.

A recombinant lentiviral vector comprising a coding sequence for an OR protein fused to a coding sequence for a fluorescent protein or a subunit of a fluorescent protein, and a promoter active in human cells operatively linked to the coding sequences. A recombinant lentivirus comprising a recombinant genome comprising an RNA that generates an ANCH sequence upon retrotranscription. A recombinant eukaryotic cell comprising a genomically integrated DNA copy of the recombinant lentiviral vector. A method of observing lentiviral DNA in a eukaryotic cell, comprising: providing a recombinant eukaryotic cell that produces a fusion protein comprising an OR protein, fused to a fluorescent protein or a subunit of a fluorescent protein; infecting the recombinant eukaryotic cell with a recombinant lentivirus comprising a recombinant genome comprising an RNA that generates an ANCH sequence upon retro-transcription, under conditions sufficient for reverse transcription of the recombinant lentiviral genome comprising an ANCH sequence; allowing the OR protein to bind to the ANCH sequence; and detecting the fluorescent protein or subunit of the fluorescent protein to thereby observe the lentiviral DNA in the eukaryotic cell. This tool can be suitable also for in vivo applications (e.g. humanized mice) as well as for screening of new antiretroviral compounds. The HIV-1 ANCHOR system can be extended to the study of other viruses or for the screening of antiviral compounds, e.g. against SARS-CoV2.