Detection method for biological activity of chicken interferon alpha according to luciferase reporter gene method and application of detection 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
CN108728514AInactive Publication Date: 2018-11-02ANHUI JIUCHUAN BIOTECH
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
ANHUI JIUCHUAN BIOTECH
Publication Date
2018-11-02
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention discloses a detection method for biological activity of chicken interferon alpha according to a luciferase reporter gene method and application of the detection method. The detection method comprises the following steps: obtaining a pMx1 gene fragment of chicken protein Mx1 through PCR amplification; inserting the pMx1 gene fragment into the 5' end of a pGL3-basic vector luc gene, and then performing PCR amplification to obtain a pMx1-luc fusion gene fragment; using the pMx1-luc fusion gene fragment to replace the pCMV and EGFP gene fragments in a pEGFP-N1 vector, constructing apMx1-luc plasmid, transfecting cells, and selecting stably transfected cell lines; performing cloning culture on the selected stably transfected cell lines; preparing a standard curve by using a chicken interferon alpha standard subjected to gradient dilution, adding a to-be-detected chicken interferon alpha sample to the cells subjected to cloning culture, for co-incubation, then detecting the fluorescence intensity, and evaluating the titer of the to-be-detected chicken interferon alpha sample according to the standard curve.
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Description

technical field

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

[0002] Interferon α 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 α in cells has been studied thoroughly, mainly through the activation of JAK-STAT signal cascade. Interferon α 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

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