马氏珠母贝TRADD基因克隆与组织表达分析

Gene Cloning and Tissue Expression Analysis of TRADD from Pinctada martensii

【目的】克隆马氏珠母贝(Pinctada martensii)肿瘤坏死因子受体相关死亡域蛋白(TRADD)基因,并分析其在各组织中的表达。【方法】利用cDNA末端快速扩增技术(RACE)克隆获得马氏珠母贝PmTRADD基因的cDNA全长序列,利用实时荧光定量PCR(qPCR)方法分析PmTRADD基因在马氏珠母贝不同组织中的表达模式。【结果与结论】PmTRADD包含5′非编码区101bp,3′非编码区144bp和开放阅读框(ORF)591bp,编码196个氨基酸。序列分析表明,PmTRADD没有信号肽和跨膜结构域,C端含有一个死亡结构域(DEATH)。将PmTRADD死亡结构域的氨基酸序列与其他物种的TRADD死亡结构域序列进行比对,发现不同物种的TRADD死亡结构域序列同源性较低。PmTRADD在马氏珠母贝各组织中均有不同程度表达,在鳃组织中表达最高,肝胰腺次之,闭壳肌中基本无表达。

【Objective】To clone the tumor necrosis factor receptor associated death domain protein (TRADD) gene and analyze its expression patterns in tissues of Pinctada martensii.【Method】The full-length cDNA sequence of PmTRADD was cloned by rapid amplification of cDNA ends (RACE) and the expression of this gene was analyzed by RT-PCR.【Result and Conclusion】The PmTRADD full-length cDNA contained 5′ UTR (101 bp), the 3′ UTR (144 bp) and open reading frame (591 bp) which encoded 196 amino acids. Analysis of deduced amino acids showed that it has no signal peptide and transmembrane domain, and the C-terminus contains a death domain (DEATH). Results from multi-sequence comparisons showed that the DEATH domain of PmTRADD has low homology compared with other species. PmTRADD gene was with higher expression level in the gill, followed by the hepatopancreas, and almost no expression in the adductor muscle.