13 results about "Membrane Protein Gene" patented technology

The invention discloses a cell membrane protein replacement type oncolytic virus vector and application thereof. The cell membrane protein replacement type oncolytic virus vector is rhabdoviridae virus, and a nucleotide sequence for coding G protein in a genome of the rhabdoviridae virus is replaced by a nucleotide sequence for coding an antibody and a nucleotide sequence for coding a spike protein truncation of coronavirus. The invention also discloses a construction method of the cell membrane protein replacement type oncolytic virus vector for expressing the antibody, the non-replicated virus vector is used for expressing the antibody sequence for the first time, and meanwhile, the novel coronavirus cell membrane protein is embedded into the virus surface, so that the cell membrane protein replacement type oncolytic virus vector can be rapidly produced in a suspension cell in a large scale; through removal of virus cell membrane protein genes, the virus cell membrane protein genes cannot be continuously replicated in vivo, so that the safety of the virus cell membrane protein genes is ensured, the tumor immunosuppression condition is improved, and an organism can be stimulated to generate a neutralizing antibody for resisting new coronavirus while tumor cells are killed.

Provided are a recombinant Escherichia coli with improved ability to secrete extracellular endoglucanase (BcsZ) and a construction method therefor. The recombinant Escherichia coli is constructed by using Escherichia coli JBZ-DH5α as a starting strain and inserting a membrane protein coding gene. The membrane protein gene is one of the following coding genes: a coding gene of zinc transporter permease znuB, a coding gene of DUF1449 family inner membrane protein ylaC, and a coding gene of UPF0410 family inner membrane protein ymgE.

The application belongs to the field of biotechnology and agricultural pest control, and particularly relates to a membrane protein Sec61 alpha gene and application of dsRNA of the membrane protein Sec61 alpha gene in pest control. After cloning and sequencing of the Sec61 alpha gene of Locusta migratoria, the sequence of the Sec61 alpha gene of SEQ ID NO:1 is obtained; then the gene fragment of SEQ ID NO:2 is selected for synthesis of double-stranded RNA (dsRNA). After the dsRNA of the gene is injected into the body cavity of the Locusta migratoria, specific Sec61 alpha genes can be silenced, and the Locusta migratoria stops feeding and successively dies before molting. The application provides a new specific molecular target for pest control based on RNA interference, and has very important application value.

The invention belongs to the field of bioengineering, and particularly relates to a construction method and application of a transgenic mouse for screening tissue cell membrane protein in vivo. Aiming at the problems of low in-vivo research flux, unstable library, limited tissue applicability and the like of the traditional membrane protein gene, sgRNA library plasmids are constructed by designing sgRNA of a membrane protein target gene and a control gene; three-positive-mouse containing Cas9, UBC-CreERT2 and sgRNA libraries is obtained through embryo microinjection and hybridization, genotype identification, inducer regulation and control, disease modeling, multi-tissue sample treatment and NGS analysis technical systems are established in a matched mode, and membrane protein gene in-vivo high-throughput screening is achieved. The model can guarantee library stability and screening reliability, covers multi-tissue research scenes, links in-vitro screening and in-vivo verification and is suitable for membrane protein function analysis and disease target mining, the technical process can be popularized to other gene families, and systematic biological research is promoted.

The present application relates to the technical field of veterinary biological products, and particularly relates to a C-type chicken infectious rhinitis subunit vaccine and a preparation method and application thereof. The present application uses Region 2 (abbreviated as TP5-BP5-BS-C-R2) of the outer membrane protein HMTp210 gene of the C-type chicken infectious rhinitis containing an immune enhancement sequence fusion of TP5-BP5-BS as an immunogen to prepare a subunit vaccine. The C-type chicken infectious rhinitis subunit vaccine prepared by the method of the present application has high safety, overcomes the side reactions such as injection site swelling caused by the whole bacteria inactivated vaccine, and can quickly induce the immune animals to produce high levels of neutralizing antibodies, realizes the efficient immune protection of the C-type chicken infectious rhinitis, and the immune effect is better than that of the C-type rhinitis subunit vaccine and the whole bacteria inactivated vaccine without the immune enhancement sequence. Moreover, the preparation process of the subunit vaccine is simple, the cost is low, and the subunit vaccine is easy to mass-produce.

The invention belongs to the field of bioengineering, and particularly relates to a construction method and application of a transgenic mouse for in-vivo screening of a T cell membrane protein gene. Aiming at the problems of low in-vivo screening research efficiency, library instability, tissue applicability limitation and the like of traditional T cell membrane protein genes, sgRNA library plasmids are constructed by designing sgRNA of a T cell membrane protein target gene and a control gene; three positive mice containing Cas9, Ubc-CreERT2 and sgRNA libraries are obtained through embryo microinjection and hybridization, genotype identification, inducer regulation and control, disease modeling, multi-tissue sample treatment and NGS analysis technical systems are established in a matched mode, and in-vivo multi-tissue sample high-throughput screening of T cell membrane protein genes is achieved. The model can guarantee library stability and screening reliability, covers multi-tissue research scenes, links in-vitro screening and in-vivo verification and is suitable for T cell membrane protein gene function analysis and disease target mining, the technical process can be popularized to other gene families, and systematic biological research is promoted.

The application discloses application of a feather needle grass unknown function domain 679 membrane protein gene SpDUF679 in plant drought stress tolerance, and belongs to the technical field of genetic engineering. The CDS sequence of the unknown function domain 679 membrane protein gene SpDUF679 is shown as SEQ ID NO. 5, and the amino acid sequence of the encoded protein is shown as SEQ ID NO. 6. The application finds a new unknown function domain membrane protein 679 of feather needle grass, names it as SpDUF679, successfully constructs a plant expression vector of the gene, and verifies that the gene has important functions of being capable of enhancing the drought stress resistance of plants in transgenic plants. The application has important significance for revealing the stress resistance mechanism of feather needle grass, enriching the theory of plant stress resistance molecular biology, and improving the stress resistance of plants.

The present invention relates to the field of biological vaccine research and development technology, and in particular, to a VSV vector-based highly pathogenic H5 subtype avian influenza inactivated vaccine and a method for preparing same. The method comprises: using a vesicular stomatitis virus with the deletion of the surface envelope protein gene as a vector, inserting the NA gene and the HA gene of an H5 subtype highly pathogenic avian influenza virus strain between the M gene and the L gene of the vector in the form of an expression cassette, and acquiring a recombinant virus by means of rescue via reverse genetic technology; inoculating the recombinant virus into susceptible cells, culturing for replication and proliferation, and then harvesting a viral solution; inactivating the viral solution to acquire an inactivated viral solution; and adding the inactivated viral solution to a pharmaceutically or veterinarily acceptable carrier, excipient, medium, or adjuvant to acquire an inactivated vaccine. The provided H5 subtype avian influenza inactivated vaccine can induce a high hemagglutination inhibition titer of avian influenza virus on day 21 after chick immunization. The provided inactivated vaccine has good safety and immunogenicity.

The application belongs to the field of biology and discloses a plasmid containing Haemophilus parasuis OmpAm outer membrane protein gene and Mycoplasma hyopneumoniae membrane protein gene; the Mycoplasma hyopneumoniae membrane protein gene is one or more combinations of Mycoplasma hyopneumoniae membrane protein P46 gene, Mycoplasma hyopneumoniae membrane protein P65 gene and Mycoplasma hyopneumoniae membrane protein P159 gene. The plasmid contains OmpAm outer membrane protein gene and Mycoplasma hyopneumoniae membrane protein gene, target genes amplified from the vector by primers are used to prepare recombinant Haemophilus parasuis by natural transformation, and one or more of Mhp antigen proteins such as P46 protein, P65 protein and P159 protein can be effectively expressed, so that the effect of double immunization is achieved. Meanwhile, the application also provides recombinant Haemophilus parasuis and application thereof.

The invention relates to a grass carp cell membrane protein gene CRISPR (clustered regularly interspaced short palindromic repeats) activated plasmid library. The library comprises CRISPR activated plasmids for targeted activation of 3498 candidate grass carp cell membrane protein genes; the invention further relates to a recombinant lentivirus capable of infecting fish cells and a packaging system of the recombinant lentivirus. The invention provides a CRISPR (clustered regularly interspaced short palindromic repeats) activation library aiming at related genes of grass carp cell membrane protein for the first time, and further provides recombinant lentivirus capable of infecting fish cells and a packaging system of the recombinant lentivirus. Protein on a grass carp cell membrane is identified through experiments, genes which exist in genome prediction but are not actually expressed can be excluded, and a library which accurately reflects related genes of grass carp cell membrane protein is formed. The CRISPR activation library provided by the invention avoids the situation of cell death after essential genes in a gene knockout library are knocked out, can comprehensively aim at essential or non-essential genes for cell survival, and can be widely applied to scenes such as virus receptor identification and / or gene function research and / or drug screening.

The application discloses a preparation method and application of a recombinant adenovirus spontaneously generating virus-like particles, and relates to the technical field of biology. The preparation method of the recombinant adenovirus comprises the following steps: connecting a virus membrane protein gene and a budding-promoting EARP gene to construct a recombinant exogenous virus membrane protein gene; inserting the recombinant exogenous virus membrane protein gene into an E3 region of an adenovirus skeleton plasmid to obtain a recombinant skeleton plasmid; inserting the recombinant exogenous virus membrane protein gene into an E1 region of an adenovirus shuttle plasmid to obtain a recombinant shuttle plasmid; and co-transfecting the recombinant skeleton plasmid and the recombinant shuttle plasmid into a host cell to obtain a recombinant adenovirus spontaneously generating virus-like particles (VLP). The recombinant adenovirus constructed by using the preparation method can spontaneously assemble VLPs containing virus outer membrane proteins in an in-vivo or in-vitro infection process, and thus the immunogenicity of a vaccine can be greatly improved, the VLP production process is simplified, and the production cost of the vaccine is reduced.

The invention belongs to the technical field of molecular biological detection, and particularly relates to a product for LAMP-CRISPR detection of porcine epidemic diarrhea virus, which comprises an LAMP primer group designed based on highly conservative membrane protein (M) gene in porcine epidemic diarrhea virus genome and a group of sgRNA. According to the product disclosed by the invention, an LAMP amplification system and CRISPR / Cas12b detection can be fused in one-tube constant-temperature reaction, only one reaction temperature needs to be set, a cover does not need to be opened in the whole process, and the operation is simple, convenient and efficient. And the product has high sensitivity and excellent specificity, effectively eliminates false positive interference, and is more suitable for clinical rapid and accurate detection. Meanwhile, the product disclosed by the invention can also be combined with an immune colloidal gold test strip to be better applied clinically, so that the detection of the porcine epidemic diarrhea virus does not depend on large-scale experimental equipment, and the obvious advantages of high efficiency and convenience are achieved.

The invention provides recombinant spirulina and application thereof in preparation of a white spot syndrome virus (WSSV) oral vaccine. The recombinant spirulina expression system is obtained by introducing an expression cassette containing a white spot syndrome virus envelope protein gene into starting spirulina to stably express the virus envelope protein. According to the present invention, the spirulina strain suitable for genetic transformation is screened, and the key structure protein of the white spot syndrome virus is efficiently expressed in the spirulina strain, such that the obtained recombinant spirulina cell can induce the shellfish such as shrimp to produce the immunocompetence against the white spot syndrome virus, and has the function of oral vaccine. Besides, the spirulina is a natural additive commonly used in crustacean feeds, so that when the recombinant spirulina is added into the feeds, the white spot syndrome virus can be effectively prevented on the premise that the cost is not obviously increased, and an economical and convenient virus prevention and control means is provided for the breeding industry of crustacean animals such as shrimps and the like.