鲤鱼TLR8基因的克隆及其组织表达谱分析

Cloning and Expression Profiling of TLR8 Gene in Common Carp (Cyprinus carpio)

为获得鲤鱼TLR8基因序列,并明确其在不同组织的表达情况,通过RT-RCR方法克隆了鲤鱼TLR8基因序列,利用荧光定量PCR分析了TLR8基因的表达特征。结果表明:所得鲤鱼TLR8基因序列1349 bp,开放阅读框(ORF)序列1113 bp,Gen Bank登录号为KT886923。另外,该克隆序列与金钱鲃、斑马鱼的序列同源性分别达到91%、82%,但与人和鼠等哺乳动物的序列同源性却很低。表达分析显示,TLR8基因在所有被检测的鲤鱼组织中均表达,尤其是在肠道和脾脏中的表达水平显著高于其他组织(P<0.05)。本研究成功获得了鲤鱼TLR8基因序列,证明其在肠道、脾脏和皮肤中的特异性表达,为进一步研究TLR8基因在鲤鱼免疫应答中的作用提供了基础。

In order to obtain sequence of TLR8 gene and its expression profiling in different tissues of common carp （Cyprinus carpio）, we cloned TLR8 and analyzed the mRNA expression profiling of TLR8 by RT-PCR. The results showed that sequence of TLR8 in C. carpio （GenBank accession No. KT886923） was 1349 bp, with an ORF of 1113 bp. In addition, the cloned TLR8 sequence in C. carpio shared 91% and 82% homologywith that in Sinocyclocheilus graham and Danio rerio, respectively, whereas shared relatively low homology with that in mammals such as Homo sapiens and Mus musculns. The expression analysis showed that TLR8 gene was extensively expressed in all tested tissues in C. carpio, with the highest expression level in gut, followed by in spleen（P〈0.05）. We successfully obtained the sequence of TLR8 in C. carpio, and demonstrated its tissue- specific expression in gut, spleen and skin, providing a basis for further investigation on the roles of TLR8 in immune response of C. carpio.