Functional analysis of Fenneropenaeus chinensis anti-lipopolysaccharide factor promoter regulated by lipopolysaccharide and (1,3)-β-D-glucan

Current knowledge on cis-regulatory elements of immune genes of shrimp is poor. In this study, the genomic sequence of the Fenneropenaeus chinensis anti-lipopolysaccharide factor (ALFFc) gene was obtained by using PCR and genome walking techniques, and the promoter was identified. The ALFFc gene contained three exons interrupted by two introns. Immune-related transcription factor binding sites recognized by nuclear factor-kappa B, octamer binding protein 1, GATA binding factor 1 and specificity protein 1 were identified in the regin from +1 to −702. The activity of ALFFc promoter was analyzed in insect sf9 cell lines. The putative promoter sequence of pALF-702 drive the expression of reporter EGFP gene successfully by adding lipopolysaccharide or (1,3)-β-D-glucan, but the shorter promoter sequence pALF-318 is only by (1,3)-β-D-glucan. The results pointed out that these transcription elements might contribute to the differences in promoter of ALFFc. Our results would provide supports for future studies to identify the functional transcription elements in the ALF promoter and to expand our knowledge on regulation of innate immune genes in Chinese shrimp.