Enhanced green fluorescent protein recombinant H5N1 subtype influenza virus and its application

Abstract

The invention discloses an enhanced green fluorescent protein recombinant H5N1 subtype influenza virus and its application. The genome of the recombinant H5N1 subtype influenza virus is composed of eight RNA fragments of the following a) and b): a) seven RNA fragments which are same as the genome of wild type H5N1 subtype influenza virus contains RNA fragment 1 encoding PB2 protein, RNA fragment 2 encoding PB1 and PB1-F2 proteins, RNA fragment 3 encoding PA protein, RNA fragment 4 encoding HA protein, RNA fragment 5 encoding NP protein, RNA fragment 7 encoding M1 and M2 proteins, RNA fragment 8 encoding NS1 and NS2 proteins; b) fragment 6' which is obtained by the insertion of RNA fragment containing green fluorescent protein coding RNA to 5'-noncoding region of RNA fragment 6 encoding NA protein in the genome of wild type H5N1 subtype influenza virus. According to the invention, rapid quantitative determination of the recombinant virus can be performed by observation of fluorescence positive cell number and fluorescence intensity, cumbersome procedure of regular titration is saved, and a convenient and effective tool for anti-H5N1 subtype influenza virus drug screening and vaccine evaluation is provided.

Description

technical field [0001] The present invention relates to a recombinant H5N1 subtype influenza virus and application thereof, in particular to a recombinant H5N1 subtype influenza virus using enhanced green fluorescent protein (EGFP) as a reporter gene and application thereof. Background technique [0002] The H5N1 subtype influenza virus is a type of influenza A virus that is mainly transmitted in birds. In recent years, reports of H5N1 subtype influenza virus infecting humans have increased year by year. As of August 2012, 608 people have been reported infected in 15 countries and regions around the world, of which 359 people died, with a mortality rate as high as 59%. Infection titer is the basic data required for virus-related basic research and drug and vaccine development. At present, the infectious titer of H5N1 subtype influenza virus is mainly determined by measuring half the tissue culture infectious dose (TCID50) or plaque forming unit (PFU) of the virus, the opera...

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Problems solved by technology

[0004] At present, there is no report on the recombinant H5N1 subtype influenza virus with enhanced green fluorescent protein (EGFP) as the reporter gene

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