313 results about "Viral genomes" patented technology

The invention relates to a micro-fluidic chip for sequencing a single virus genome and a sequencing method. The micro-fluidic chip comprises a first functional area, a delay pipeline, a fusion pipeline, a second functional area and a fusion electrode, the first functional area is used for generating first liquid drops of virus-in-oil and lysate; two ends of the delay pipeline are respectively connected with the first functional area and the fusion pipeline; the second functional area is used for generating second liquid drops of the oil bag amplification reaction liquid, and the second functional area is connected with the fusion pipeline to enable the second liquid drops to enter the fusion pipeline; a fusion point is arranged on the downstream of the joint of the fusion pipeline and the second functional area, and the fusion electrode can act on the fusion point to fuse the first liquid drop and the second liquid drop. According to the micro-fluidic chip disclosed by the invention, single-virus whole genome amplification can be completely completed on a single chip, so that the operation steps are simplified, the complexity of sample treatment is reduced, pollution and sample loss are effectively avoided, and the efficiency, the stability and the reliability are improved.

The invention provides a viral genome splicing method and system combining a reference sequence and de novo splicing, and the method comprises the steps: carrying out the quality control of the sequencing data of a viral genome sequence, and removing a host genome sequence in the sequencing data based on a k-mers strategy; splicing the sequencing data from the beginning to generate a preliminary assembling sequence of the viral genome; performing virus classification annotation on the virus genome preliminary assembly sequence based on a blastn tool, and selecting a highly similar virus genome sequence from a virus library as a reference sequence according to the virus classification annotation; generating a data set according to the reference sequence and the virus genome preliminary assembly sequence, dividing a plurality of sequence clusters according to the similarity between the sequences in the data set, and generating a consensus sequence; comparing the reads after the host is removed with a consensus sequence, and adjusting the consensus sequence according to a comparison result to obtain a final virus genome sequence. According to the method, the accuracy of virus genome splicing is improved, and the universality is high.

The invention discloses a chimeric PEDV (porcine epidemic diarrhea virus) infectious clone plasmid, a PEDV chimeric virus as well as a construction method and application thereof. According to the invention, a PEDV genome is used as a skeleton for the first time, the chimeric virus for expressing the extracellular domain of the TGEV S protein is successfully rescued, and it is found that the TCID50 of the chimeric virus for expressing the extracellular domain of the TGEV S protein can be increased by 6.2 times compared with that of the chimeric PEDV for expressing the TGEV S protein. The recombinant virus is used as an inactivated vaccine for immunizing mice, so that the immunogenicity of TGEV S protein can be greatly improved, a body is stimulated to generate a higher-level neutralizing antibody aiming at TGEV, and the titer of the neutralizing antibody aiming at TGEV is improved by 16.8 times. Therefore, more possibilities are provided for modification of viral genomes, a novel method for improving immunogenicity of the TGEV inactivated vaccine is invented, and a basis is provided for research and development of the TGEV vaccine.

The invention provides a gene extraction method for AAV third-generation sequencing, and particularly provides a simple and efficient rAAV gene extraction method for third-generation sequencing, which comprises the following steps: firstly, performing shell breaking on rAAV purified virus by using protease K to obtain an initial reaction solution; performing alkali denaturation on the initial reaction solution, neutralizing, purifying by using a DNA product purification kit, and eluting by using nuclease-free water to obtain denatured virus genome DNA; and finally, incubating and annealing the denatured virus genome DNA to finally form the complete double-stranded DNA. Compared with a traditional AAV genome extraction method, the method provided by the invention is simpler and more efficient, and the extracted DNA is higher in integrity.

Oncolytic viral vectors that incorporate one or more of the following features: viral replication restriction by insertion of microRNA (miRNA) target sequences into the viral genome; disruption of oncogenic miRNA function; cancer microenvironment remodeling; and cancer cell targeting by incorporation of protease-activated antibodies into the viral particle. Such viral vectors can be used for the treatment and prevention of cancer.

The invention relates to the technical field of agricultural biology, and particularly provides a dsRNA nanocrystallization preparation for preventing and treating areca yellows as well as a preparation method and application of the dsRNA nanocrystallization preparation. Areca yellows are a destructive disease caused by areca symptomless virus type 1 (APV1), eight key target gene segments of an APV1 virus genome are screened, specific double-stranded RNA is designed, a nano-carrier is used for wrapping and delivering a dsRNA mixture obtained by transcription of the eight target gene segments, and the areca yellows are obtained by screening. And a nanocrystallization preparation for improving the environmental stability and RNA interference efficiency of the dsRNA is prepared. Experimental results show that the nanocrystallization preparation can effectively inhibit the key gene expression of APV1 virus, reduce the virus load and improve the resistance of areca catechu to yellows. The invention provides an accurate, efficient and environment-friendly biological prevention and control strategy, and has a wide agricultural application prospect.

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.

The invention provides a primer group, a kit and a method for targeted amplification of a type I dengue virus genome, and application of the primer group, the kit and the method, relates to dengue virus detection, and belongs to the technical field of genome detection. The universal primer group for amplifying the I-type dengue virus genome is designed, targeted amplification can be performed on the I-type dengue virus genome, sequencing and sequence splicing are performed by utilizing a three-generation sequencing platform after a library is constructed, and the I-type dengue virus genome is obtained. The method provided by the invention can amplify all I-type dengue virus subtypes, and has the advantages of large flux, low cost and simple sequence splicing.

The present invention relates to methods for producing a preparation comprising recombinant AAV (rAAV), such methods comprising a step of mechanical lysis on mammalian producer cells, and methods for increasing the viral genome titre and / or capsid titre of a preparation comprising recombinant adeno-associated virus, related uses, and preparations obtained by or obtainable by the methods.

The invention belongs to the technical field of biological detection, and relates to a method for detecting lentivirus titer by a double-standard-curve RT-qPCR (Reverse Transcription-Quantitative Polymerase Chain Reaction) method. Infecting the target cells by adopting a to-be-detected sample containing a recombinant lentivirus, and collecting and extracting genome DNA of the infected target cells; carrying out a viral genome RT-qPCR amplification reaction and an internal reference genome RT-qPCR amplification reaction by respectively taking the original virus standard substance and the internal reference standard substance which are diluted in a gradient manner and the extracted target cell genome DNA as templates; and calculating the lentivirus titer corresponding to the sample according to data obtained by the RT-qPCR amplification reaction. The method can be used for absolute quantitative qPCR detection of lentivirus titer, can accurately determine lentivirus successfully integrated into a cell genome, and reflects the real infection ability of the virus; in addition, the technology has high sensitivity and specificity, is not influenced by the extraction efficiency of the cell genome DNA, and ensures the reliability of the detection result.

The invention belongs to the field of medicines, and particularly relates to application of fructose-1, 6-diphosphate in preparation of a medicine for preventing and treating herpes simplex virus type 1 infection. The fructose-1, 6-diphosphate (FBP) can be used for inhibiting replication of RNA (Ribonucleic Acid) and DNA (Deoxyribose Nucleic Acid) viruses in vitro and in vivo. Experiments show that the inhibition effect does not depend on I-type interferon-mediated innate immunity to a great extent, but occurs through HMGB1 (High Mobility Group Box 1). After virus infection, FBP reduces the protein level of HMGB1 by promoting lysosome degradation of HMGB1. In addition, the FBP can directly inhibit the combination of the HMGB1 and the viral genome, so that the virus replication is inhibited. The key effect of FBP in inhibition of virus replication is highlighted through research, and a potential target spot for controlling virus infection is provided.

The invention discloses a recombinant rabies virus genome plasmid for high-sensitivity expression of red fluorescent protein, a recombinant rabies virus as well as a preparation method and application of the recombinant rabies virus. The preparation method specifically comprises the following steps: (1) constructing the recombinant rabies virus genome plasmid for high-sensitivity expression of red fluorescent protein as shown in SEQ ID NO.2; (2) preparing a recombinant rabies virus for expressing the red fluorescent protein with high sensitivity; and (3) application of the recombinant rabies virus for high-sensitivity expression of the red fluorescent protein in neural circuit tracing. The recombinant rabies virus capable of highly sensitively expressing the red fluorescent protein can efficiently mark an upstream neural network in a retrograde manner, so that the problem of low efficiency of expressing red fluorescence in the existing method is solved; the method has wide application value and market prospect in the aspects of neural circuit structure and function analysis, neuron sparse marking, vaccine and drug research and development, disease model establishment, neural network loss and reconstruction detection, virus replication or pathogenic mechanism research and the like.

The present disclosure relates to the field of molecular virology, including nucleic acid molecules comprising a modified viral genome or self-replicating RNA, pharmaceutical compositions containing said nucleic acid molecules, and the use of such nucleic acid molecules and compositions for producing a desired product in a cell culture or in vivo. Also provided are methods for eliciting a pharmacodynamic effect in a subject in need thereof, as well as methods for preventing and / or treating a variety of health conditions.

In some aspects, cardiac-specific expression cassettes are provided herein. In some aspects, provided herein is an expression cassette comprising a polynucleotide sequence encoding a gene product for therapy of a heart disease, wherein the polynucleotide sequence is operably linked to promoter (e.g., a cardiac-specific promoter), and optionally an enhancer (e.g., a cardiac-specific enhancer). In some aspects, the disclosure provides recombinant adeno-associated virus (rAAV) virions, comprising a capsid protein and a viral genome comprising an expression cassette comprising a polynucleotide sequence encoding a therapeutic gene product, e.g., dwarf open reading frame (DWORF) polypeptide, operably linked to a promoter, the expression cassette flanked by inverted terminal repeats. The disclosure further provides pharmaceutical compositions and methods of treating or preventing heart disease.

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.

Disclosed herein are prokaryotic homologs of viperin (pVips), and nucleotide and nucleoside analogs produced from pVips. These nucleotide and nucleoside analogs stop nucleotide chain synthesis and provide host cells with resistance to viral infections by targeting actively replicating viral genome. Further, these nucleotide and nucleoside analogs decrease DNA replication in malignant cells. Further disclosed are methods of identifying pVips, and nucleotide and nucleoside analogs produced thereof.

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.

The invention discloses a different-length genome sequence coding method based on improved self-attention and Mamba double modules, and relates to a different-length genome sequence coding method. The invention aims to solve the problem that a genome sequence cannot be reasonably input into a neural network model in the prior art. The method comprises the following steps: in a data processing sequence feature extraction process, extracting kmer position feature information, kmer centrality feature information and inter-kmer relevance feature information of genome sequences with different lengths; in the pre-training process, a model based on improved self-attention and Mamba double modules is constructed, a sequence coding model can capture universal features of genome sequences through mask pre-training, and universal representation vectors of the same dimension of the genome sequences of different lengths are obtained; in the fine tuning process, by using specific biological task data for fine tuning training, the coding model can be better matched with various virus-based genomes such as virus recognition, virus host and virus lifestyle recognition. The invention belongs to the technical field of deep learning and gene engineering.

Methods of characterizing biomolecular formulations are provided. In some examples, there is provided a method of characterizing a viral particle formulation, the method comprising subjecting the viral particle formulation to analytical ultracentrifugation under sedimentation velocity conditions, measuring sedimentation of the viral particle at a plurality of points in time, determining a friction ratio function c (s, ff0) and a sedimentation coefficient for the viral particle formulation, mapping the sedimentation coefficient (S) with the c (s, ff0) to obtain c (s, ff0), and characterizing the c (s, ff0). Ff0) distribution, and characterizing the presence and / or relative composition of viral particles comprising a totiviral genome, a partial viral genome, an over-packaged viral genome, and / or an empty viral particle in the viral particle formulation.

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 belongs to the field of biological medicine, and particularly relates to a recombinant adenovirus and application thereof. The technical problem to be solved by the invention is to provide the recombinant adenovirus with good tumor inhibition effect and high efficiency. The technical scheme of the invention is a recombinant adenovirus. The recombinant adenovirus is a replication-defective adenovirus, and an exogenous nucleic acid fragment is operably inserted into a viral genome; the exogenous nucleic acid fragment is a coding gene of human p53 protein, a coding gene of GM-CSF and a coding gene of RGD. The coding gene of the human p53 protein, the coding gene of the GM-CSF and the coding gene of the RGD are jointly constructed into the replication-deficient adenovirus, so that the adenovirus with multiple advantages is obtained, the tumor cell killing efficiency is improved by 2-4 times compared with that before recombination, and a new effective choice is provided for tumor immunotherapy.

The invention belongs to the technical field of virus detection, and particularly relates to an anguilla positive reovirus (Anguilla positive reovirus) AORV-1025 as well as a primer group, a kit and a method for detecting the anguilla positive reovirus. According to the invention, a conserved sequence of an RdRp gene coded by an L3 segment of an eel positive reovirus genome is taken as a target, a primer group consisting of a primer pair and a probe is designed, and the primer group is used for detecting several different positive reoviruses including the eel positive reovirus and viruses possibly existing in a detection sample. The detection result is high in sensitivity, strong in specificity and excellent in repeatability, the eel positive reovirus in a sample can be rapidly and accurately detected in a fluorescent quantitative mode, the method is applied to research on eel positive reovirus infection, and the method has important significance on pathogen monitoring, differential diagnosis and guide prevention and control of eel viral diseases.

The invention provides systems and methods for analyzing viruses by representing viral genetic diversity with a directed acyclic graph (DAG), which allows genetic sequencing technology to detect rare variations and represent otherwise difficult-to-document diversity within a sample. Additionally, a host-specific sequence DAG can be used to effectively segregate viral nucleic acid sequence reads from host sequence reads when a sample from a host is subject to sequencing. Known viral genomes can be represented using a viral reference DAG and the viral sequence reads from the sample can be compared to viral DAG to identify viral species or strains from which the reads were derived. Where the viral sequence reads indicate great genetic diversity in the virus that was infecting the host, those reads can be assembled into a DAG that itself properly represents that diversity.

The invention relates to the technical field of gene engineering, in particular to a gene vector based on CsCMV mediation and a preparation method and application thereof. On the basis of a pCsCMV-CM vector, a Bsm BI site is successfully inserted into a non-coding region (UTR) behind a termination codon (TAG) at a CsCMV CP 3'end through an In-Fusion seamless cloning method, an infectious clone pCsCMV-Bs vector based on a Golden gate Coning strategy is constructed, a target gene segment can be rapidly cloned into a virus genome with high throughput, and the target gene segment can be cloned into the virus genome. The method is applied to cassava gene silencing and gene editing, the stability is higher, the phenotype duration time is longer, and a new genetic tool is provided for rapid and efficient gene function verification and directional character improvement of cassava.

The present disclosure generally relates to methods for generating functional self-replicating RNA from non-functional positive single-stranded RNA (+ssRNA) viral genomes or srRNA, such as alphavirus genomes. More specifically, the methods relate to the assembly of multiple nucleic acid fragments to generate functional srRNA de novo. Also provided are compositions, nucleic acid constructs, vectors and recombinant cells comprising such functional srRNA. The present disclosure further discloses methods for inducing a pharmacodynamic effect in a subject, as well as methods for preventing and / or treating a health condition in a subject in need thereof.

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 present disclosure relates to the field of molecular virology, including nucleic acid molecules comprising modified viral genomes or self-replicating RNAs, pharmaceutical compositions containing the same, and the use of such nucleic acid molecules and compositions for production of desired products in cell cultures or in a living body. Also provided are methods for eliciting a pharmacodynamic effect in a subject in need thereof, as well as methods for preventing and / or treating various health conditions.

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.