Pseudo-virus and packaging method thereof and drug evaluation system

Abstract

The invention relates to pseudo-virus. The pseudo-virus comprises a plasmid for expressing envelope protein Spike, a reporter gene plasmid and a packaging helper plasmid, wherein the plasmid for expressing the envelope protein Spike contains a CBA promoter and a human optimized Spike expression sequence, and is used for driving the expression of the envelope protein Spike. The pseudo-virus disclosed by the invention can simulate the process of infecting cells by wild novel coronavirus, can be used for researching the relationship between virus and host cells, cloning a receptor of virus and replacing a traditional neutralization test, and is used for researching differential diagnosis of diseases, evaluation and screening of antiviral drugs, evaluation of vaccine immune effect and the like. The invention also relates to a packaging method of the pseudo-virus and a novel coronavirus virus drug evaluation system containing the pseudo-virus, wherein the pseudo-virus is used for infectingthe 293T monoclonal cell which overexpresses the human ACE2 gene under the condition of drug interference, and by detecting the expression quantity of a reporter gene in the pseudo-virus (such as fluorescence intensity detection), the effectiveness of the drug is preliminarily evaluated.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a pseudovirus, a packaging method thereof and a drug evaluation system. Background technique [0002] In the study of virus function, the most direct and efficient method is of course to manipulate wild-type live viruses. However, for highly pathogenic and infectious viruses such as the new coronavirus, the operation of wild-type live viruses needs to be carried out in biosafety protection (P3) laboratories above level 3. At present, there are only more than 40 P3 laboratories in China, and most of them Belongs to the CDC system, non-conventional scientific research purposes. In addition, in order to prevent biological hazards, the P3 laboratory has very strict protection conditions, which greatly reduces the operating efficiency. [0003] In order to be able to conduct research on the new coronavirus in the second-level biosafety laboratory, pseudovirus technology is a very ...

AI Technical Summary

Problems solved by technology

However, in 2004, when scientists packaged the SARS-CoV pseudovirus system, they found that in the packaging cells, the CMV promoter could not drive the expression of wild-type S protein. Through codon optimization, the amount of S protein increased, but Still can't make up for the lack of promoters

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