Detection method for biological activity of porcine interferon alpha through luciferase reporter gene method
The invention discloses a detection method for the biological activity of a porcine interferon alpha through a luciferase reporter gene method and application of the detection method. The method comprises the following steps: obtaining a gene segment of pMx1 of porcine Mx1 protein by adopting PCR (P...
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Main Authors | , , , , , , |
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Format | Patent |
Language | Chinese English |
Published |
26.10.2018
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Subjects | |
Online Access | Get full text |
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Summary: | The invention discloses a detection method for the biological activity of a porcine interferon alpha through a luciferase reporter gene method and application of the detection method. The method comprises the following steps: obtaining a gene segment of pMx1 of porcine 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 culture on the screened stably-transfected cell line; preparing a standard curve by taking a porcine interferon alpha standard product which is subjected to gradient dilution; adding a porcine interferon alpha sample to be det |
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Bibliography: | Application Number: CN20181641986 |