71 results about "Polyhedrin" patented technology

The invention relates to a porcine circovirus type 2 subunit vaccine, and a preparation method and an application thereof. The recombinant baculovirus contains double promoters (a polyhedrin protein promoter and a P10 promoter), and can express double copies of Cap protein coding genes, such that protein expression efficiency is substantially improved. Also, Cap protein expressed by an inserted exogenous gene does not contain excess sequences, such that virus-like particles (VLPs) can be effectively formed, expressed protein immunogenicity is improved, and the content of produced antigen is high. According to the porcine circovirus type 2 subunit vaccine provided by the invention, protein yield and quality of porcine circovirus type 2 subunit vaccine are substantially improved, and prepared vaccine compositions have the advantages of stable and long-lasting immune effect, high safety, and the like.

Cypoviruses and baculoviruses are notoriously difficult to eradicate because the virus particles are embedded in micron-sized protein crystals called polyhedra. The remarkable stability of polyhedra means that like bacterial spores these insect viruses remain infectious for years in soil. Although these unique in vivo protein crystals have been extensively characterized since the early 1900s, their atomic organization remains elusive. Here we describe the 2 crystal structure of both recombinant and infectious silkworm cypovirus polyhedra determined using 5-12 micron crystals purified from insect cells. These are the smallest crystals yet used for de novo X-ray protein structure determination. It was found that polyhedra are made of trimers of the viral polyhedrin protein and contain nucleotides. Although the shape of these building blocks is reminiscent of some capsid trimers, polyhedrin has a new fold and has evolved to assemble in vivo into 3-D cubic crystals rather than icosahedral shells. The polyhedrin trimers are extensively cross-linked in polyhedra by non-covalent interactions and pack with an exquisite molecular complementarity similar to that of antigen-antibody complexes. The resulting ultra-stable and sealed crystals shield the virus particles from environmental damage. The structure suggests that polyhedra can serve as the basis for the development of robust and versatile nanoparticles for biotechnological applications such as in cell culture systems, microarrays and biopesticides.

Insect-derived promoters for foreign proteins expression in insect cells. Regulatory polynucleotide sequences which drive the expression of major insect proteins (hexamerins) at specific evolution stages of larva have been isolated from insects (Trichoplusia ni) in the present invention. Said regulatory polynucleotide sequences promote stronger foreign gene expression in the baculovirus system than the conventional polyhedrin promoter. Additionally, the combination of the new larva-derived promoters of the invention with the pL promoter increased baculovirus expression levels up to 61%-375% (depending on the time of infection) with respect to conventional baculo viruses used in the biotechnology industry. Promoter pB2 also drives gene expression at earlier times than polyhedrin promoter, being this feature a great advantage for correct protein folding and posttranslational modification.

Provided is an insect control sheet using a polyhedrin protein polyhedron (Cry polyhedron) in which the insecticidal protein (Cry toxin) produced by Bacillus thuringiensis is fixed. The insect control sheet contains the Cry polyhedron, and is used by floating on water. The insect control sheet floats on the surface of water, and is obtained by layering a pure matrix layer (20) and a toxin-containing matrix layer (30) containing a Cry polyhedron (51) onto the undersurface of a sheet-shaped first sheet substrate (10). The pure matrix layer (20) comprises an easily decomposable or water-soluble second material, while the toxin-containing matrix layer (30) comprises the Cry polyhedron and an easily decomposable or water-soluble third material. The Cry polyhedron is gradually released from the toxin-containing matrix layer into the water on which the control sheet is floating.

An insect control sheet containing a Cry polyhedron prepared by fixing an insecticidal protein (a Cry toxin) produced by Bacillus thuringiensis to a polyhedron of polyhedrin protein is provided. The insect control sheet contains the Cry polyhedron and is used by floating on water. The insect control sheet is floatable on water, and includes a pure matrix layer 20 and a toxin-containing matrix layer 30 containing the Cry polyhedron 51 which are layered on the underside of a sheet-shaped first sheet substrate 10. The pure matrix layer 20 is composed of a degradable or water-soluble second material and the toxin-containing matrix layer 30 is composed of a degradable or water-soluble third material and the Cry polyhedron. The toxin-containing matrix layer sustainably releases the Cry polyhedron to the water on which the insect control sheet is floated.

The invention relates to an animal genetic engineering interferon compound preparation, the production method and the clinical application. The animal genetic engineering interferon compound preparation is prepared by extracting the animal peripheral blood or spleen lymphocytes, culturing, inducing by in vitro inductor, extracting the cell total RNA by Trizol, designing the specific primer, cloning the interferon gene by RT-PCR technology and connecting to the pGEM-T carrier by T-A strategy; the primer is designed again, the both ends are added with different restriction endonucleases digestion sequences, initiation codon ATG and termination codon TAA, leading peptide sequences are removed, the PCR amplification is carried out by taking the recombination T carrier as the template, so as to obtain the expression fragment of the animal interferon gene; the expression fragment is carried out with the rare codon mutation, and gel recovery target fragment is connected on the carrier of pFastBacDual with the same double restriction endonucleases digestion after double restriction endonucleases digestion; the interferon-Alpha of an animal is cloned to multiple cloning sites under the control of Polyhedrin promoter, the interferon-Gamma of the same species animal is cloned to multiple cloning sites under the control of p10 promoter; the constructed carrier of pFastBacDual plus interferon Alpha plus interferon Gamma are transfected to DH10BacE.coli to carry out recombination; the constructed recombinant bacmid is extracted and purified after the blue white screening and the PCR identification, and the transfection of SF9 insect cell is carried out by a liposome method; the expression product is carried out with identification and purification; the animal genetic engineering interferon compound preparation is carried out with anti-virus activity detection and the evaluation of the clinical application effects.