Molecular cloning and expression of allograft inflammatory factor 1 in Haliotis diversicolor under stresses

Abstract

Allograft inflammatory factor-1(AIF-1)is a kind of calcium-binding protein with EF-hand domain.AIF-1 mRNA is robustly induced by IFN-γ in murine macrophages.It is involved in transplant rejection,immune inflammatory reaction,non-inflammatory injury and so on.In this study,a molluscan AIF-1 gene,HdAIF-1,was cloned for the first time from Haliotis diversicolor.Its full-length cDNA sequence is 942 bp,with a 456 bp open reading frame encoding a protein of 151 aa.Quantitative real-time PCR results indicated that HdAIF-1 could be detected in all examined tissues,with the highest level in hemolymph and gill.Under thermal stress,HdAIF-1 was up-regulated in gill significantly at temperature-rise period and achieved the highest level when up to 31 ℃.However,the expression level of HdAIF-1 in hemolymph and hepatopancreas did not show significant difference between thermal group and control group from the first phase to the fourth,and its expression level was up-regulated significantly at 96 h in these two tissues.Under hypoxia stress,the expression level of HdAIF-1 in hemolymph showed no significant difference between control and exposed groups.However,it was down-regulated at 24 h and up-regulated at 192 h remarkably in gill.After Vibrio parahaemolyticus challenge,the HdAIF-1 expression level achieved the highest level at 3 h in hemolymph,reduced to the control level at 12 h,and then up-regulated significantly at both 24 h and 48 h.These results suggest that HdAIF-1 may play important roles as an immune factor under different stresses.