Abstract

Secretory carrier membrane proteins (SCAMPs) are widely distributed integral membrane proteins implicated in membrane trafficking. Secretory carrier membrane protein 5 (SCAMP5) is expected to be involved in regulation of the immune response because it is expressed in a variety of immune tissues and promotes the secretion of cytokines in monocytes and macrophages. In this study, we performed an analysis of the molecular characteristics and phylogenetic of the SCAMP5 gene identified in Pagrus major (PmSCAMP5). In addition, we analysed PmSCAMP5 gene expression levels in the tissues of red sea bream infected with various pathogens [Edwardsiella piscicida (E. piscicida), Streptococcus iniae (S. iniae) and Red sea bream iridovirus (RSIV)], and we analysed PmSCAMP5 gene expression levels in the tissues of healthy red sea bream. This study was carried out to provide basic data on the non-specific immune system of the red sea bream.

Highlights

  • Data on molecular characterization and gene expression analysis of secretory carrierassociated membrane protein 5 (SCAMP5) from the red sea bream (Pagrus major)

  • Secretory carrier membrane protein 5 (SCAMP5) is expected to be involved in regulation of the immune response because it is expressed in a variety of immune tissues and promotes the secretion of cytokines in monocytes and macrophages

  • We performed an analysis of the molecular characteristics and phylogenetic of the SCAMP5 gene identified in Pagrus major (PmSCAMP5)

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Summary

Molecular characterization and phylogenetic analysis

An ORF sequence of the PmSCAMP5 gene was obtained from the liver of red sea bream stimulated with lipopolysaccharide (LPS) through Generation Sequencing (NGS) analysis. The amino acid sequence of PmSCAMP5 was predicted using the GENETYX ver. Development, Japan) and the BLASTX program of the National Center for Biotechnology Information (NCBI). The molecular weight and isoelectric point were predicted using the ProtParam tool of ExPASy Proteomics Serve, and specific domain was identified using Simple Modular Architecture Research Too. The multiple alignment of the PmSCAMP5 amino acid sequence and the SCAMP5 amino acid sequence of other species registered in peptide sequence database of NCBI was analysed using ClustalW analysis. The phylogenetic analysis of PmSCAMP5 was performed using the neighbour-joining (NJ) method of MEGA 4 program and bootstrap sampling was repeated 2,000 times [1]

Experimental animal and microbes
Infection experiment
Total RNA extraction and reverse transcription
RT-qPCR analysis of PmSCAMP5 and statistical analysis
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