Anguilla japonica antibacterial peptide Cathelicidin1 gene promoter and application thereof

Abstract

The invention relates to an anguilla japonica antibacterial peptide Cathelicidin1 gene promoter and an application thereof. According to the anguilla japonica antibacterial peptide Cathelicidin1 gene promoter, the anguilla japonica antibacterial peptide Cathelicidin1 gene promoter sequence is successfully cloned, and the anguilla japonica antibacterial peptide Cathelicidin1 gene promoter pGL3-Cathelicidin1-pro luciferase reporter plasmid is successfully constructed; and reporter gene analysis experiments prove that the anguilla japonica antibacterial peptide Cathelicidin1 gene promoter can be induced and activated by a virus simulant poly I:C, an important surface antigen LPS of gram-negative bacteria and an important pathogenic bacterium aeromonas hydrophila of aquatic animals. The invention discloses clone of an anguilla japonica antibacterial peptide Cathelicidin1 gene promoter and inducible expression analysis of strong promoter activity of the anguilla japonica antibacterial peptide Cathelicidin1 gene promoter, and provides a good experimental system for researching an expression regulation mechanism of a fish antibacterial peptide Cathelicidin1 gene, a natural immune response mechanism of fish for resisting pathogenic bacterium infection, and particularly a regulation mechanism of a fish inflammation-related NF-[kappa]B and MAPK signal path network, and therefore, conditions are created for constructing an expression vector by using the promoter to efficiently express an exogenous gene or applying the promoter to transgenic fish construction.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a Japanese eel antibacterial peptide Cathelicidin1 gene promoter and application thereof. Background technique [0002] Antimicrobial peptides (AMPs) are a class of biologically active small molecule peptides, which are an important part of the body's innate immune defense system and can be effective against Gram-negative bacteria, Gram-positive bacteria, fungi, viruses and parasitic Both have good inhibitory or killing effects, and play an important role in natural immune response[Chung, C.-R., J.-H.Jhong, Z.Wang, W.Chen, Wan, Horng and T. - Y. Lee (2020). "Characterization and Identification of Natural Antimicrobial Peptides on Different Organisms." International Journal of Molecular Sciences 21: 986.]. Fish, like mammals and other higher vertebrates, have both innate and adaptive immune systems. As an important part of the innate immune system of fish, antimicrob...

AI Technical Summary

Problems solved by technology

The promoter is the key factor determining gene expression and regulation. In order to study the expression regulation mechanism of the fish Cathelicidin1 gene, we obtained the possible 5′ flank regulation of the Japanese eel Cathelicidin1 gene by comparing the open reading frame sequence of the Japanese eel Cathelicidin1 gene with the genome sequence The promoter sequence of the Japanese eel Cathelicidin1 gene was obtained after primer design and PCR cloning verification, and the analysis showed that the promoter had the transcription binding sites of C / EBPalp, NF-kappaB, GATA, GAAA, and AP1 in the reported rainbow trout Cathelicidin1 promoter However, CREBP and TFIID transcription binding sites are missing, and there are unique transcription factor binding sites such as RAP1, Sp1, AP-2alph, RAP1, USF, NF-EM5, etc., showing the species of the Japanese eel Cathelicidin1 gene promoter sequence specificity

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