ORF016 gene deletion type goat pox virus strain and preparation method and application thereof

• Main Authors: TANG NA, SHEN ZHIQIANG, ZHANG QIAN, WANG JINLIANG, WEI FENG, REN YANLING, YUAN WENXIAN
• Format: Patent
• Language: Chinese; English
• Published: 30.10.2020
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; HUMAN NECESSITIES; HYGIENE; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; MEDICAL OR VETERINARY SCIENCE; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; ORGANIC CHEMISTRY; PEPTIDES; PHYSICS; PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS ORMEDICINAL PREPARATIONS; SPIRITS; TESTING; VINEGAR; WINE

The invention discloses gRNA, a recombinant plasmid, an ORF016 gene deletion type goat pox virus strain and a preparation method and application of the ORF016 gene deletion type goat pox virus strain,and the sequence of the gRNA is shown as SEQ ID NO: 1 or SEQ ID NO: 2. According to the invention, the ORF016 gene deletion type goat pox virus strain is obtained by a gene knockout method for the first time, the CRISPR technology is applied to the goat pox virus for the first time, meanwhile, the ORF016 gene is knocked out through a reverse transfection method, and the knockout efficiency of themethod reaches 31.6% and is far higher than that in the prior art. Meanwhile, the SDLVDUORF016 strain obtained by the invention is a strain only lacking ORF016, the deletion of the gene has no influence on the growth characteristics of the strain, and the strain can be used for goat pox vaccines, detection reagents, goat pox virus identification and the like. 本发明公开了一种gRNA，所述gRNA序列如SEQ ID NO：1或SEQ ID NO：2所示，本发明还公开了重组质粒、ORF