仿刺参C1q基因的克隆与表达分析  

Cloning and mRNA Expression Analysis of C1q Gene in Sea Cucumber Apostichopus japonicus

作　　者：陈仲[1] CHEN Zhong(Liaoning Key Laboratory of Marine Fishery Molecular Biology,Liaoning Ocean and Fisheries Science Research Institute,Dalian 116023,China)

机构地区：[1]辽宁省海洋水产科学研究院,辽宁省海洋水产动物种质改良与优良苗种繁育重点实验室,辽宁大连116023

出　　处：《水产科学》2023年第6期933-944,共12页Fisheries Science

基　　金：辽宁省百千万人才工程资助项目(2021921071);辽宁省农业科学院基本科研业务费计划项目(2022XTCX0504);辽宁省农业科学院学科建设计划项目(2022DD268143);大连市重点领域创新团队支持计划项目(2021RT08)。

摘　　要：C1q作为补体经典激活途径的识别蛋白,对于机体的免疫作用至关重要。为探明C1q基因在仿刺参免疫防御中的作用,利用构建的仿刺参cDNA文库,通过RACE技术克隆得到仿刺参C1q(AjC1q)的全长cDNA,其序列全长为1717 bp,其中5′-端非翻译区为110 bp,3′-端非翻译区为698 bp,开放阅读框为909 bp,编码302个氨基酸。AjC1q蛋白具有球状C1q结构域,为三聚体,与自然界中大多数动物中的C1q结构域有着较高的同源性。实时荧光定量PCR检测结果显示:在仿刺参从受精卵到幼参的发育过程中,AjC1q基因的mRNA均有表达,并且在樽形期的表达水平明显增加,到五触手幼体期达到峰值,表明补体系统可能在樽形期被广泛激活;AjC1q基因的mRNA在不同组织中均有表达,其中在管足中的表达水平较高;不同病原菌刺激后,AjC1q基因的转录表达均在较短时间内发生上调,表明在病原菌刺激下AjC1q基因能够快速表达。AjC1q可能在仿刺参发育过程中以及免疫应答中起着重要作用。C1q,as the complement recognition protein of the classical activation pathway,is very important for the immunity of the body.In order to study the role of C1q gene in the immune defense of sea cucumber Apostichopus japonicus,the full-length cDNA sequence of sea cucumber C1q(AjC1q)cDNA was cloned by the homologous gene cloning and RACE,and other molecular biology techniques.The full-length cDNA sequence was 1717 bp,in which the 5'-terminal untranslated region was 110 bp,the 3-terminal untranslated region was 698 bp,and the open reading frame was 909 bp,encoding 302 amino acids.AjC1q protein has a globular C1q domain and is a trimer,which has high homology with the C1q domain of most animals in wild.The mRNA of AjC1q was expressed in the whole development of sea cucumber.The expression increased significantly in the doliolaria stage and reached the peak in the pentactula stage,indicating that the complement system may be widely activated in the doliolaria stage.And the mRNA of AjC1q was expressed in different tissues and was highly expressed in tube feet.In addition,the AjC1q gene could be up-regulated in a short time after stimulation by different pathogens,indicating that AjC1q is rapidly expressed under the stimulation of pathogens.So AjC1q may play an important role in the development of sea cucumber and in the immune response.

关 键 词：仿刺参 C1Q CDNA克隆 生物信息学 

分 类 号：Q78[生物学—分子生物学]