摘要
髓样分化因子(My D88)是TOLL样受体介导的信号通路中的一个关键接头分子,通过激活核转录因子(nuclear factor-kappa B,NF-κB)而参与机体的先天免疫。克隆了松江鲈的My D88基因(命名为Tf My D88),并对该基因进行了生物信息学和表达模式分析。结果显示,Tf My D88 c DNA序列全长1 555 bp,5'UTR长89 bp,3'UTR长599 bp;开放阅读框长度为867 bp,编码288个氨基酸。SMART软件预测Tf My D88分子的N端为一个保守的死亡结构域(death domain,DD),C端存在典型的TIR(Toll/interleukin-1 receptor)结构域。Tf My D88与其它脊椎动物My D88的氨基酸相似性达57.58%-82.64%,系统进化树分析表明Tf My D88与同属鲈形总目的花鲈和鳜鱼聚在一起,所有鱼类My D88聚为一支。Real-time PCR检测显示Tf My D88广泛表达于松江鲈各组织,但在鳃中的相对表达量最高;其次为脾脏和皮肤。LPS(lipopolysaccharide)刺激后,Tf My D88在松江鲈的血液、肝脏、皮肤、脾脏均出现明显上调。刺激2 h后,在血液Tf My D88表达量升高了近60倍,在皮肤中的表达量也升高了27倍。上述结果表明Tf My D88可能参与松江鲈先天免疫。
Myeloid differentiation factor 88（TfMyD88）is a key adaptor protein in the Toll-madiated signaling passway. In this study, we cloned the full-length cDNA from the roughskin soulpin, Trachidermus fasciatus. The full-length of MyD88 was 1 555 bp, which contained an open reading frame（ORF）of 867 bp encoding a polypeptide of 288 amino acids. The deduced amino acid sequence has a conserved death domain（DD）at the N-terminal and a typical Toll/interleukin-1 receptor（TIR）domain at the C-terminal. The mRNA expression patterns of TfMyD88 in healthy and LPS challenged roughskin soulpin were detected using quantitative Real-time PCR. MyD88 was expressed broadly in the blood, heart, liver, gill, intestine, skin, musle, kidney, spleen, brain, with the highest expression in the gill. The expression of TfMyD88 post challenge with LPS（lipopolysaccharide）was detected in blood, liver, gill, skin and spleen. The up-regulation of the expression levels of TfMyD88 in all the detccted tissues after chanllaged by LPS suggests that MyD88 plays an important role in roughskin soulpin defenses against bacterial infection.
出处
《生物技术通报》
CAS
CSCD
北大核心
2015年第2期135-142,共8页
Biotechnology Bulletin
基金
威海市科技攻关计划项目(0000413420608)
威海市科委项目(1070432121313)
