Detection method for biological activity of sheep interferon tau through luciferase reporter gene method

A biological activity and reporter gene technology, applied in the field of interferon activity detection, can solve the problems of biological safety hazards, poor repeatability, and low accuracy of live viruses, so as to avoid biological safety hazards, simplify the detection process, and shorten the detection time. the effect of time
CN108707643AInactive Publication Date: 2018-10-26WUHU YINGTE FEIER BIOLOGICAL PROD IND RES INST CO LTD
1 Cites 0 Cited by

Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
WUHU YINGTE FEIER BIOLOGICAL PROD IND RES INST CO LTD
Publication Date
2018-10-26
Estimated Expiration
Not applicable · inactive patent

Smart Images

  • Figure 1
    Figure 1
  • Figure 2
    Figure 2
  • Figure 3
    Figure 3
Patent Text Reader

Abstract

The invention discloses a detection method for the biological activity of a sheep interferon tau through a luciferase reporter gene method and application of the detection method. The method comprisesthe following steps: obtaining a gene segment of pMx1 of sheep Mx1 protein by adopting PCR (Polymerase Chain Reaction) amplification; inserting the gene segment of the pMx1 into a 5' end of a pGL3-basic vector luc gene; then carrying out the PCR amplification to obtain a pMx1-luc fused gene segment; replacing gene segments of pCMV (Porcine Cytomegalovirus) and EGFP (Enhanced Green Fluorescent Protein) in a pEGFP-N1 vector by utilizing the pMx1-luc fused gene segment; constructing a pMx1-luc plasmid and transfecting a cell; selecting a stably-transfected cell line; carrying out clonal cultureon the screened stably-transfected cell line; preparing a standard curve by taking a sheep interferon tau standard product which is subjected to gradient dilution; adding a sheep interferon tau sampleto be detected into cells subjected to the clonal culture for co-incubating; then detecting the fluorescence intensity and combining the standard curve to evaluate the valence of the sheep interferontau sample to be detected.
Need to check novelty before this filing date? Find Prior Art

Description

technical field

[0001] The invention relates to a method for detecting the biological activity of sheep interferon tau by a luciferase reporter gene method, and belongs to the technical field of interferon activity detection. Background technique

[0002] Interferon τ is widely used in the treatment of diseases in antiviral infection and immune dysfunction. As a cytokine, it can activate a series of intracellular signal transduction pathways by binding to specific receptors indicated by target cells, showing antiviral effects. or immunomodulatory effects. At present, the signal transduction pathway of interferon tau in cells has been studied thoroughly, mainly through the activation of JAK-STAT signal cascade. Interferon tau binds to receptors to activate receptor-related JAK1 and TYK2, phosphorylate STAT1 and STAT2 tyrosine, phosphorylated STAT1 and STAT2 form dimers and transfer into the nucleus, and assemble interferon regulatory factor 9 (IRF9) to form 3-mer interferon...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More