245 results about "Polerovirus" patented technology

Disclosed and claimed is a new insect cell line, Sf900+, ATCC CRL-12579. The insect cell line was established from Lepidoptera, Noctuidae, Spodoptera frugiperda Sf-9 (ATCC CRL-1711) through multiple rounds of limiting dilution and selection in a serum-free insect medium supplemented with added human insulin. The insect cell line is useful in BEVS or as an adjuvant and has many characteristics and advantages. Also disclosed and claimed are recombinant proteins from recombinant baculovirus expression in insect cells such as Sf900+ cells, for instance, HA, NA, EPO, CD4, CEA, and thrombospondin.

The invention mainly aims to provide a porcine circovirus type 2 (PCV2) subunit vaccine and a preparation method thereof. Particularly, a baculovirus vector expression system is utilized to express a large amount of recombinant open reading frame type 2 (ORF2) protein in insect cells, so that the subunit vaccine with good immunity effect is developed. A bac-to-bac baculovirus expression system is adopted to perform whole gene amplification on the porcine circovirus type 2 ORF2 gene, and a melittin signal peptide nucleotide sequence is introduced into a terminal 5', so that the recombinant baculovirus of an open reading frame containing the melittin signal peptide nucleotide sequence and a PVC2ORF2 gene sequence is established, wherein the infected insect cell expresses the recombinant ORF2 protein with efficient and high PCV2 antigenicity; and thus the subunit vaccine containing the PCV2 recombinant ORF2 protein is prepared. The inoculation experiments of piglets show that the subunit vaccine has a good immunity protection effect.

The invention relates to a coxsackie virus A16 type virus-like particle vaccine. The inventor fortuitously finds that proteins which have better spatial configurations and are suitably cut can be obtained by expressing P1 protein of CVA16 and 3CD protein of CVA16 by infecting insect cells with rhabdovirus. The proteins can be automatically assembled into a virus-like particle which has high immunogenicity.

The invention discloses a SARS vaccine and preparing method, which comprises the following steps: 1) constructing external baculoviral surface display carrier of S protein of SARS coronary virus; 2) transmitting the carrier into susceptive cell with baculoviral genome plasmid Bacmid; obtaining recombinant baculoviral genome plasmid; 3) using the plasmid to infect insect cell; purifying the recombinant plasmid; obtaining the product.

The invention discloses a genetic engineering subunit vaccine for a porcine circovirus as well as a preparation method and application of the genetic engineering subunit vaccine. By cloning the nucleocapsid protein of the novel porcine circovirus 3 (PCV3), a PCV-Cap protein with higher purity is successfully expressed by using an escherichia coli or baculovirus expression system. The subunit vaccine for the PCV3 is successfully developed for the first time by using the PCV-Cap protein; the prepared vaccine is high in antigen purity, good in safety and strong in immunogenicity, and has no pathogenicity to pigs and other animals; the antigen has good solubility in a neutral PH buffer solution; furthermore, the preparation method is simple and low in cost, thus being suitable for large-scaleindustrial production; an effective and powerful means is provided for the prevention and control of novel PCV, and the genetic engineering subunit vaccine has a wide application prospect in the fieldof the prevention and control of the PCV3.

The invention provides rabbit hemorrhagic disease virus-like particles. The rabbit hemorrhagic disease virus-like particles are prepared by optimizing rabbit hemorrhagic disease virus VP60 gene according to insect cell biased codons, connecting the optimized VP60 gene with an insect cell efficient expression vector, and then producing the rabbit hemorrhagic disease virus-like particles through a rhabdovirus/ insect cell expression system. The rabbit hemorrhagic disease virus-like particles can be mixed with a preservative and adjuvants to prepare a rabbit hemorrhagic disease vaccine. The rabbit hemorrhagic disease virus-like particles can greatly increase the expression quantity of the rabbit hemorrhagic disease virus-like particles, and have the advantages of high safety, high immunogenicity, low cost and the like, and is easy to ferment.

The recombination, expression and application of Chinese prawn's antibacterial peptide gene are disclosed. Its recombination expression method includes configuring recombinant carrier (pET series, pGEX-4T series, pPIC series, pAO815, and baculovirus expression carrier), transferring the carrier to colibacillus and yeast and transfecting insect cells, screening engineered bacterial strain or cell, and inducing expression. The expressed product can be used to prepare feed additive, antistaling agent, cosmetics and medical products.

The present invention provides an insect infection method for use in the production of a protein with a baculovirus expression vector in the insect, the method comprising the steps of:

• • (g) providing a plurality of insect larvae or pupae;
  • (h) providing a solution comprising a wild type baculovirus or a baculovirus expression vector having a desired gene encoding a protein;
  • (i) stressing the insect larvae or pupae;
  • (j) soaking the insect larvae or pupae in the solution for an appropriate time so that they are infected with the wild type baculovirus or the baculovirus expression vector; and
  • (k) incubating the infected larvae or pupae for production of the protein; and
  • (l) harvesting the protein.

The method can treat a great quantity of larvae simultaneously, achieve batch infection of larvae or pupae, save manpower and effectively infect larvae or pupae at a high infection rate.

The invention discloses a recombinant baculovirus vector, a virus like particle, a preparation method and use and aims at providing tandem expression and accurate cutting expression of a plurality of genes with a single promoter and successfully packaging the virus like particle. The recombinant baculovirus vector is simple to operate, high in expression quantity and high in cutting efficiency. The obtained virus like particle is good in immunogenicity and does not have an exogenous sequence. The technical essential is that picornaviridae 3C/3CD gene and P1/P1op gene are expressed under the single promoter, the 3C/3CD gene and the P1/P1op gene are connected by a 2A sequence to build a 3C/3CD-2A-P1/P1op expression frame, then the 3C/3CD gene is placed under the promoter of baculovirus vector plasmid to obtain a recombinant shuttle vector, a host cell is transfected to obtain recombinant baculovirus, and the host cell is infected to obtain the virus like particle.

The invention discloses a foot and mouth disease virus recombinant virus sample particle as well as a preparation method and application thereof. The recombinant virus sample particle is commonly assembled and expressed by components in a DNA molecule composition. The DNA molecule composition contains an O-type foot and mouth disease VP0 gene, an O-type foot and mouth disease VP1 gene and an O-type foot and mouth disease VP3 gene and further contains a green fluorescent protein gene. By virtue of the property that FMDV VLPs is self-assembled through VP0, VP1 and VP3, a construction method of a baculovirus recombinant vector is improved, a green fluorescent protein label is added into the vector, and FMDV VLPs is successfully prepared by virtue of a pFBDM Bac-to-Bac system, so that a theoretical foundation is laid for the further development of safe and efficient O-type FMDV genetic engineering vaccines.

The invention relates to a porcin circovirus type 2 (PCV2) recombinant autographa californica multicapsid nucleopolyhedrovirus construction method and a subunit vaccine preparation method thereof. The novel recombinant lucerne fork vein noctuid nuclear polyhedrosis virus construction method comprises the steps that: the main surface antigen which expresses the PCV2 is used to prepare the subunit vaccine with better immunity. Specifically, in the carrier expression system of the PCV2 recombinant autographa californica multicapsid nucleopolyhedrovirus, a hepatitis E virus open reading frame (ORF2) gene of the PCV2 is directionally inserted, so the gene efficiently expresses in insect cells and has good immunogenicity, the prepared corresponding subunit vaccine can stimulate pigs to generate the protective immunoreaction against the attack of the strong virus of the PCV2.

This invention relates to the methods of detecting Penaeus monodon baculovirus (MBV). Two methods are established: the first one is a polymerase chain reaction (PCR) and the second one is an ELISA. For the PCR method, two sets of primers are designed. The first set of primers is designed from the conserved sequences of nuclear polyhedrosis viruses (NPVs) DNA polymerase genes. The second set of primers is designed from the genomic DNA of MBV. The antibody for ELISA is an antiserum against the occlusion bodies of MBV.

The invention discloses a method for producing human collagen type II, and belongs to the technical field of gene engineering. The method is characterized in that a baculovirus multi-gene expression system is adopted, insect cells are infected by the bacteria converting the baculovirus multi-gene expression system to produce baculovirus with multiple genes expressed, the virus liquid is infected in the insect cells or injected into silkworm larva bodies to produce recombinant human collagen type II full-length protein, the protein structure is that 3 peptide chains with (G-X-Y) n repetitive sequences form a three-strand helical structure, to both keep the conformation of the natural collagen type II and have the activity of the natural collagen type II. By adopting the method disclosed by the invention, multiple genes can be expressed to facilitate the production of then collagen type II full-length protein; the method disclosed by the invention is simple in process method, simple and convenient to operate and short in process route.

A method for constructing a recombinant double-expression silkworm polyhedral baculiform virus which contains the SOD gene comprises that the general RNA of the silkworm is extracted and an MnSOD primer is designed; a single-stranded cDNA is synthesized and an MnSOD gene is synthesized; the MnSOD gene is connected with a p FastBacHTa to transform XL-Blue cells; and the recombinant donor plasmid p FastBacHTa/MnSOD can be extracted; the baculiform virus DNA of the silkworm is taken as the template to synthesize the silkworm polyhedral gene and the gene is cloned to the p FastBac Dual plasmid; the MnSOD gene is inserted under the P10 promoter of the double-expression plasmid to construct the double-expression vector: Bm polyhedrin pFB dual/MnSOD; after the transinfection of the Bm polyhedrin pFB dual/MnSOD, the double-expression virus DNA which contains the MnSOD gene and the polyhedral gene is distilled; the DNA is transduced into the silkworm culture cell of transinfection with the mediation of transinfection reagent lipidosome to obtain the recombinant virus. Virus particles which can feed and infect the silkworm is provided and the object gene MnSOD is expressed by the silkworm and large-scale and industrial production is realized.

The invention discloses a preparation method and system of recombinant adeno-associated virus (rAAV), and recombinant bacmid. The method comprises: (1) separately preparing shuttle plasmids and a corresponding recombinant bacmid containing baculovirus genome; (2) integrating a rAAV core expression element which has a heterologous functional gene fragment with other expression cassettes which produce functional protein components necessary for rAAV so as to obtain a recombinant bacmid containing recombinant baculovirus genome producing the rAAV; and (3) transfecting the obtained recombinant bacmid into a host cell line for culturing. The system comprises the shuttle plasmids and the corresponding recombinant bacmid containing the baculovirus genome. The recombinant bacmid comprises at leastone expression cassette that produces functional protein components necessary for rAAV. The system has flexibility, compatibility, and higher passage stability.