Main cis-acting element of shrimp white spot syndrome virus (WSSV) iel promoter and transcription factor combined with same and application

Abstract

The invention discloses a main cis-acting element of a shrimp white spot syndrome virus (WSSV) iel promoter and a transcription factor combined with the same and application. In the invention, by starting from transcriptional regulation of WSSV iel and carrying out structural and functional analysis on the promoter of the WSSV iel through deletion and mutation, a 12-bp DNA is found to be the maincis-acting element of the WSSV iel and is a crucial factor for the high expression of the iel. In the invention, a DNA affinity chromatography method is used for purifying a protein combined with a DNA segment from a nucleus protein Sf9, the protein is identified to be PHB2 (Poly-Beta-Hydroxybutyrate 2) through biological mass spectrometry, and the interaction between the protein and the DNA segment is proved to be specific by an electrophoretic mobility shift assay. Experiment results prove that PHB2 serves as a transcription factor and is specifically combined with a 12-bp DNA sequence of the iel promoter to start WSSV immediate early gene transcription so as to further regulate the replication of the WSSV, and can be used as an effective action target of medicaments for screening medicaments for resisting the shrimp WSSV.

Description

technical field [0001] The invention relates to a main cis-acting element of a viral gene promoter and a transcription factor combined with it, in particular to the main cis-acting element of a shrimp white spot syndrome virus (WSSV) ie1 promoter and a transcription factor combined with it. Their application, the invention belongs to the field of expression and regulation of shrimp white spot syndrome virus gene. Background technique [0002] Shrimp white spot syndrome (WSS) is the most serious infectious disease to the shrimp industry. The pathogen is white spot syndrome virus (WSSV). The disease first broke out in Taiwan, China in the 1990s, and then spread rapidly to the main shrimp farming areas throughout Asia, Europe and the United States. The virus has strong infectivity and replication ability, and the host mortality rate can reach 100%. It can infect various tissues of prawns and has a wide range of hosts. It can infect prawns, crayfish, crabs, and freshwater crust...

AI Technical Summary

Problems solved by technology

The complete sequencing of the genomes of three strains of WSSV in 2001-2002 opened the prelude to the study of WSSV molecular virology, but the analysis found that most of its open reading frames had no homology with the existing gene sequences in GenBank, so it is very difficult to Difficult to study gene expression and replication patterns of other viruses

[0004] Although some progress has been made in the molecular biology research of WSSV, the most important thing is to focus on the structural protein of the virus; there is no cell line suitable for the growth of the virus, and there is no system for the molecular mechanism of virus infection and replication. There is still a lack of understanding of the mechanism of WSSV infection, especially the regulation mechanism of virus replication, and the interaction between the virus and the host; in addition, because shrimp is an invertebrate without a specific immune system, it cannot produce antibodies, so it cannot be immunized with vaccines ways to prevent the disease

So so far, there is still no effective method for the prevention and treatment of this viral disease.

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