Cloning of BTN1A1 Genes from Symphysodon aequifasciatus and Analysis of Its Expression under Aeromonas hydrophila Stress
[Objective]Butyrophilin 1A1(BTN1A1)gene from Symphysodon aequifasciatus was cloned and its express pattern under pathological infection was analyzed,to provide basis for better understanding of the function of BTN1A1 gene in S.aequifasciatu.[Method]The structure and evolutionary characteristics of two BTN1A1 genes(BTN1A1-1 and BTN1A1-2)were analyzed by bioinformatics analysis.Based on the transcriptome data of the skin of the S.aequifasciatu obtained at early stage,the CDS regions of BTN1A1-1 and BTN1A1-2 genes were used to design the primers for gene cloning.The expressions of BTN1A1-1 gene in various tissues and its expression pattern under the stimulus of Aeromonas hydrophila were analyzed by qRT-PCR.[Result]The ORF regions of BTN1A1-1 and BTN1A1-2 were 894 and 1 275 bp,encoding 298 and 424 amino acids,respectively.Both BTN1A1-1 and BTN1A1-2 had a classical structure of BTN,which contained a signal peptide,an Ig domain,an lg_like domain and a transmembrane domain.BTN1A1-2 also had a cytoplasmic domain.BTN1A1-1 showed a closer relationship with the genes found in cichlid fish.It shared the highest homology of 94.14％with the corresponding amino acid sequences of Archocentrus centrarchus;while BTN1A1-2 showed lower similarity with other types of fish,forming a special branch.The expressions of BTN1A1-1 and BTN1A1-2 were highly in gill,intestine and skin of S.aequifasciatu.The expression of BTN1A1-1 decreased significantly and then increased after the infection of A.hydrophila,while the expression of BTN1A1-2 increased significantly and then decreased.[Conclusion]The BTN1A1-1 gene was more evolutionarily conservative than BTN1A1-2.The differential expressions of BTN1A1-1 and BTN1A1-2 in response to immune stimulation suggests that they may be functionally differentiated in the immune defence of S.aequifasciatus.
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