马氏珠母贝接头蛋白CIKS的基因克隆与组织表达分析

Gene Cloning and Tissue Expression Analysis of Adapter Protein CIKS from Pinctada fucata martensii

【目的】探究马氏珠母贝接头蛋白CIKS(PmCIKS)在马氏珠母贝免疫反应中的作用。【方法】采用cDNA末端快速扩增(RACE)技术获得PmCIKS基因cDNA全长序列,运用生物信息学手段分析该序列,用实时荧光定量PCR(RT-PCR)技术检测PmCIKS基因在马氏珠母贝7个组织中的表达模式。【结果】PmCIKS基因cDNA全长为1 987 bp,其中5′UTR长为228 bp,3′UTR长为148 bp,包含24 bp的ploy A,开放阅读框(ORF)为1 611 bp,编码536个氨基酸,预测其分子质量约为61.3 ku,等电点为7.01。物种间CIKS有较高的保守性。PmCIKS在马氏珠母贝肝胰腺、性腺、闭壳肌、鳃、血细胞、外套膜和足中均有表达,其中在血细胞中表达量最高。

【Objective】To study the role of PmCIKS in the immune response of Pinctada fucata martensii,【Method】The full-length cDNA sequence of PmCIKS was obtained by rapid amplification of cDNA ends(RACE).The expression patterns of PmCIKS gene in 7 tissues were further tested by Quantitative Real-Time PCR technology.【Results】The PmCIKS full-length cDNA was 1987 bp.The 5′UTR was 228 bp in length,the 3′UTR was 148 bp,which contained 24 bp of ployA.The open reading frame(ORF)was 1611 bp,encoding 536 amino acids.Its molecular weight was predicted to be 61.3 ku and the isoelectric point was 7.01.Results from multi-sequence comparisons showed that the CIKS among species was highly conserved;RT-PCR analysis showed that PmCIKS gene was expressed in hepatopancreas,gonad,adductor muscle,gill,blood cells,mantle,and foot of P.fucata martensii with the highest expression level in blood cells.The results of this study showed that PmCIKS gene may play an important role in various biological process,especially in immune defense response of P.fucata martensii.