Sheep interferon Tau biological activity detection method and application thereof

• Main Authors: LI YASEN, XU GAOTAO, JIANG MINZHI, SHAN XUEQIN, LIU JIALU, HE ZHIYUAN, ZHAO YU, FAN YUFANG
• Format: Patent
• Language: Chinese; English
• Published: 06.11.2018
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PHYSICS; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; TESTING; VINEGAR; WINE

The invention discloses a sheep interferon Tau biological activity detection method and application thereof. The method includes following steps: adopting PCR amplification to acquire a gene segment of Mxp of sheep Mx protein; removing pCMV of pEGFP-N1 carrier plasmid; using the gene segment of Mxp of the sheep Mx protein obtained through PCR amplification to replace pCMV in original pEGFP-N1 carrier plasmid through T4DNA ligase to build pEGFP-N1-Mxp plasmid; using the pEGFP-N1-Mxp plasmid to transfect cells, and screening out a stable transfection cell line through neomycin; performing cloning culture on the screened stable transfection cell line, and adding sheep interferon Tau to co-incubate with the stable transfection cell line after going through cloning culture. In this way, Mx genepromoter activity can be activated to promote expression of EGFP in cells, and intensity of fluorescence emitted by the cells after being irradiated by an excitation light source is in positive correlation with biological ac