Cell line for detecting I type interferon biological activity and application thereof

A biological activity and interferon technology, which is applied in the field of detecting the biological activity of animal type I interferon, can solve the problems of affecting versatility, narrow application range, low sensitivity and accuracy, etc., and achieves convenient high-throughput detection. , the effect of saving testing time

Inactive Publication Date: 2009-12-16
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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AI Technical Summary

Problems solved by technology

AVA has its limitations, low sensitivity and accuracy, and has certain biological safety risks, so researchers continue to explore new methods to replace AVA
2. ELISA [1，2] Although the detection of IFN activity is convenient and quick, it can only detect the antigenicity of IFN. Some IFN-like proteins without biological activity will affect the detection results, so it is not reliable, and different types of IFN require different monoclonal antibodies, so also affects its versatility
3. Other biological methods based on the characteristics of IFN, such as cell proliferation inhibition, cell activity regulation, cell differentiation and immune regulation, all have limitations to varying degrees, or the methods are cumbersome, or the scope of application is narrow

Method used

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  • Cell line for detecting I type interferon biological activity and application thereof
  • Cell line for detecting I type interferon biological activity and application thereof
  • Cell line for detecting I type interferon biological activity and application thereof

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Experimental program
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Embodiment Construction

[0022] Interferon is an important antiviral substance, and the quantitative detection of its biological activity has important basic research and production application significance. The Mx promoter is one of the main effector transcriptional promoters of interferon, which expresses Mx protein under the action of type I IFN.

[0023] In one embodiment, the present invention clones the chicken Mx promoter (Mxp), the sequence of which is different from the reported ones, with a difference of 6 bases. A luciferase (luciferase, luc) reporter gene was connected downstream to construct the chicken type I interferon activity detection plasmid pMxp-luc. The DF-1 chicken embryo fibroblast cell line was transiently transfected with pMxp-luc, and the cells were treated with chicken IFN-α / β 24 hours later. As a result, the luciferase gene was transcribed and expressed under the action of the Mx promoter; luciferase The expression level of interferon has a good linear correlation with the...

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Abstract

The invention relates to a method for detecting I type interferon biological activity, in particular to a method for quantitatively detecting I type interferon biological activity of animals, such as chicken, cow and pig, based on Mx promoter and luciferase report gene. Compared with a reported detection method based on Mx promoter-report gene, the method needs shorter time, thereby greatly saving detection time; compared with a traditional antivirus quantitative detection method, the method has the advantages of fast detection, convenience, accuracy, sensitivity, convenience for high-throughput detection, and the like, and has important application value in research of poultry and even mammal interferon biological base and application and industrialization process.

Description

technical field [0001] The invention relates to a method for detecting the biological activity of animal type I interferon. More specifically, the present invention relates to a method for quantitatively detecting type I interferon biological activity in animals based on chicken Mx promoter and luciferase reporter gene. Background technique [0002] Interferon (interferon, IFN) is an important antiviral active substance and an important part of the body's non-specific immune system. It has been widely used in animal and human anti-infection and clinical treatment of tumors. However, for a long time, there has been a lack of an ideal standard quantitative detection method for IFN activity. Traditional IFN activity quantitative means mainly include: 1. Antiviral assays (antiviral assays, AVA), commonly used viruses are vesicular stomatitis virus (Vesicular stomatitis virus, VSV) and encephalomyocarditis virus (Encephalomyocarditis virus, EMCV) [1] . AVA has its limitations,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/02C12Q1/66C12N5/10
Inventor 步志高陈伟业
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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