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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Format | Patent |
Language | Chinese English |
Published |
26.10.2018
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Abstract | 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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AbstractList | 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 |
Author | LI YASEN XU GAOTAO SHAN XUEQIN ZHAO YU JIANG MINZHI LIU JIALU FAN YUFANG |
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DocumentTitleAlternate | 荧光素酶报告基因法猪干扰素α生物学活性检测方法 |
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Snippet | The invention discloses a detection method for the biological activity of a porcine interferon alpha through a luciferase reporter gene method and application... |
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SubjectTerms | 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 |
Title | Detection method for biological activity of porcine interferon alpha through luciferase reporter gene method |
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