Cloning of X-linked inhibitor of apoptosis protein XIAP gene and its response to stimulation in sea urchin Strongylocentrotus intermedius challenged with pathogenic microorganisms
LIU Shengmei, SHANG Fengqin, CHEN Yadong, CHANG Yaqing,WANG Xiuli, QIU Xuemei, LIU Yang*
Key Laboratory of Mariculture & Stock Enhancement in North China’s Sea, Ministry of Agriculture and Rural Affairs, Dalian Ocean University, Dalian 116023, China
Abstract: To investigate the role of X-linked inhibitor of apoptosis protein (XIAP) gene in the natural immune system of sea urchin Strongylocentrotusintermedius, the cDNA sequence of XIAP gene was cloned using rapid amplification of cDNA ends (RACE) technique, and distribution in various tissues including tube foot, peristomial membrane, intestine, and gonad and the expression of XIAP gene were analyzed in coelomocyte in the sea urchin exposed to stimulation including lipopolysaccharide (LPS), peptidoglycan (PGN), polyinosinic-polycytidylic acid (Poly I:C), Vibriofortis and whole glucan particles (WGP) by real-time quantitative PCR. The results showed that the length of XIAP cDNA was 4 894 bp, with 5′ UTR of 459 bp, the open reading frame of 2 331 bp and the 3′ UTR of 2 104 bp, encoding a total of 776 amino acids with three BIR structural domains and one RING finger structural domain, with a predicted relative molecular mass of 86 960 and an isoelectric point of 5.96, as acidic protein. XIAP was expressed in all tissues examined, significantly higher expression in the ovary than that in the tube foot, coelomocyte, peristomial membrane, intestine and testis (P<0.05), after stimulation with PGN, LPS, Poly I:C, WGP and V.fortis. XIAP gene expression was up-regulated in coelomocyte significantly (P<0.05), with the maximal values 12 h after PGN, LPS and Poly I:C stimulation, and with the maximal values 24 h after WGP stimulation and 6 h after V.fortis stimulation. The findings indicated that the clonally obtained XIAP gene was involved in the immune response of the sea urchin and played an important role in its natural immunity.
刘圣美, 尚凤芹, 陈亚东, 常亚青, 王秀利, 仇雪梅, 刘洋. 虾夷马粪海胆X染色体连锁凋亡抑制蛋白XIAP基因克隆及不同病原微生物刺激后的表达响应[J]. 大连海洋大学学报, 2022, 37(4): 627-635.
LIU Shengmei, SHANG Fengqin, CHEN Yadong, CHANG Yaqing, WANG Xiuli, QIU Xuemei, LIU Yang. Cloning of X-linked inhibitor of apoptosis protein XIAP gene and its response to stimulation in sea urchin Strongylocentrotus intermedius challenged with pathogenic microorganisms. Journal of Dalian Ocean University, 2022, 37(4): 627-635.