摘要
TLR13（Toll like receptor 13）是一种TLR模式识别受体,属于TLR11家族,在机体抵抗细菌、真菌、病毒和寄生虫的感染过程起重要的作用。为研究马氏珠母贝TLR13（PmTLR13）在马氏珠母贝免疫反应中的作用,本研究采用RACE扩增技术获得了PmTLR13基因cDNA全长序列,并且运用实时荧光定量PCR（Real-time PCR）技术检测PmTLR13基因在马氏珠母贝7个组织中的表达模式。结果显示,Pm TLR13基因序列全长2 750 bp,其中5＇UTR长为25 bp,3＇UTR长为34 bp,开放阅读框（ORF）为2 106 bp,编码701个氨基酸,预测其分子量约为81.22 kD,等电点为8.77。SMART分析结果显示PmTLR13具有富含亮氨酸的重复序列（LRRs）、TIR域、跨膜域和信号肽,符合典型的TLR家族特征;多序列比对结果表明物种间TLR13具有较高的保守性;RT-PCR数据分析表明,PmTLR13基因在马氏珠母贝肝胰腺、性腺、闭壳肌、鳃、血细胞、中央膜区、边缘膜区中均有表达,其中鳃中表达量最高,其次是外套膜边缘区和肝胰腺。研究结果表明,PmTLR13可能在马氏珠母贝免疫防御反应中担任着重要的角色。
TLR13（Toll like receptor 13） is a kind of TLR pattern recognition receptors, belonging to the family of TLR11, which plays an important role in resisting bacteria, fungi, viruses and parasites infection in the process. In order to explore the role of immune response of Pm TLR13 in Pinctada fucata martensii, we used c DNA End Rapid Amplification（ RACE） cloning technology and got the c DNA full length of Pm TLR13 from Pinctada fucata martensii, testing i ts expression patterns in seven tissues by Real-Time PCR technology. Results showed that the total length of Pm TLR13 gene was 2 750 bp, including a 5＇ UTR of 25 bp, a 3＇ UTR of 34 bp and an open reading frame（ORF） of 2 106 bp which encoded 701 amino acids. The predicted molecular weight was 81.22 k D, and isoelectric point was 8.77. SMART software analysis showed that Pm TLR13 had Leucine rich repeats（LRRs）, TIR domain, transmembrane domain and the signal peptide, conforming to the typical characteristics of TLR family.Multiple sequence alignment indicated that TLR13 was higher conservative among species. Real-time PCR data revealed that Pm TLR13 was expressed in seven tissues, including hepatopancreas, gonads, adductor muscle, gil l,hemocytes, mantle central, mantle edge, with the highest expression in gill, followed by mantle edge and hepatopancreas. The research results showed that Pm TLR13 might play an important role in Pinctada fucata martensii in immune defense reaction.
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2018年第1期238-246,共9页
Genomics and Applied Biology
基金
国家自然科学基金“马氏珠母贝植核育珠移植耐受免疫相关基因与蛋白质鉴定及关联度研究”(31472306)
广东省科技计划项目“海水珍珠贝功能基因的开发与应用研究”(2012A031100010)
广东省海港建设与渔业产业发展专项“马氏珠母贝抗菌功能多肽的提取与应用研究”(A201608B15)共同资助
