Construction method of PRV gE/gI dual-gene deletion strains for expressing ASFV P30 protein

• Main Authors: ZHU LING, XU ZHIWEN, ZENG YUBING
• Format: Patent
• Language: Chinese; English
• Published: 31.07.2020
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; HUMAN NECESSITIES; HYGIENE; MEDICAL OR VETERINARY SCIENCE; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS ORMEDICINAL PREPARATIONS; SPIRITS; VINEGAR; WINE

The invention discloses a construction method of PRV gE/gI dual-gene deletion strains for expressing ASFV P30 protein and belongs to the biopharmaceutical field. The construction method specially comprises the following steps of S1, preparing a reagent; S2, preparing cloned plasmids; S3, constructing a PRV eukaryon transfer plasmid pEGFP-CP204L; S4, performing cotransfection on DNA of PRV and pEGFP-CP204L plasmids. P30 protein gene (CP204L) fragments are synthesized, the transfer plasmid pEGFP-CP204L is constructed, a classical homologous recombination manner is adopted for inserting an ASFV CP204L gene into a PRV genome to replace gI and gE virulence genes, and recombinant pseudorabies virus strains rXJgE/gI-CP204L for constructing co-expression P30 protein can be used for preventing ASFVand PRV infection, and the cloned plasmids are simple to prepare, low in cost and suitable for industrial production. 本发明公开了生物医药领域的一种表达ASFV P30蛋白的PRV gE/gI双基因缺失株的构建方法，具体包括以下步骤，S1：试剂准备；S2：克隆质粒的制备；S3：构建PRV真核转移质粒pEGFP-CP204L；S4