翘嘴鳜雄性特异NLRP3基因cDNA的克隆及表达分析  

Cloning and Expression Analysis of Male-specific NLRP3 Gene cDNA from Siniperca chuatsi

作　　者：欧阳号锋 韩崇 卢灏铭 朱巧莹 马泽昊 蒲海琦 李水生[1] 张勇[1] OUYANG Haofeng;HAN Chong;LU Haoming;ZHU Qiaoying;MA Zehao;PU Haiqi;LI Shuisheng;ZHANG Yong(Institute of Aquatic Economic Animals&Guangdong Provincial Key Laboratory for Aquatic Economic Animals,Sun Yat-sen University,Guangzhou 510275,China)

机构地区：[1]中山大学生命科学学院水生经济动物研究所暨广东省水生经济动物良种繁育重点实验室,广州510275

出　　处：《海南热带海洋学院学报》2021年第5期1-8,共8页Journal of Hainan Tropical Ocean University

基　　金：国家重点研发计划项目(2018YFD0901203);广东基础与应用基础研究基金项目(2019B1515120072);广州市科学与技术规划项目(201904020043)。

摘　　要：NLRP3基因是能够识别PAMPs和胞内危险信号的1种重要的炎性小体,参与机体对体内外病原微生物的防御和维持自身稳态。首次发现翘嘴鳜存在1个雄性特异的NLRP3基因。通过设计基因特异扩增引物,并在22条雌鱼和22条雄鱼中进行PCR扩增,证实了该基因只在翘嘴鳜雄鱼中存在。为了解该NLRP3基因的潜在功能,成功克隆得到该基因的全长蛋白编码序列,并通过序列比对和系统发育分析,初步证明该基因具有NACHT、LRR和PYD等多个关键结构域,在鲈形目鱼类中具有较高的保守性。另外,通过荧光定量分析的方法研究了NLRP3基因在雄性翘嘴鳜不同组织以及在不同时期性腺中的表达差异,发现翘嘴鳜NLRP3基因在肝脏、胃和肠等组织有较高的表达,且在翘嘴鳜雄性性别分化早期有显著高表达,在性别分化后几乎不表达。NLRP3 gene is an important inflammasome which can identify PAMPs and intracellular danger signals,and thus is involved in the body’s defense,in vivo and in vitro,against pathogens as well as the maintenance of homeostasis.In the current study,a male-specific NLRP3 gene was found,for the first time,in Siniperca chuatsi.By designing specific gene amplification primers and PCR amplification in 22 female and 22 male fish,it was confirmed that the gene only exists in male fish.In order to understand the potential function of the NLRP3 gene,the full-length coding sequence of NLRP3 gene was successfully cloned in the present study.Through sequence alignment and phylogenetic analysis,it was preliminary proved that NACHT,LRR and PYD are the key domains of NLRP3 gene,and that the gene is highly conserved in Perciformes.In addition,by fluorescence quantitative analysis,examined was the expression of NLRP3 gene in male different tissues and in the gonads during different periods.It was found that NLRP3 gene has a high expression in the liver,stomach and intestinal tissue,and a significantly high expression in the early male sexual differentiation in contrast to almost no expression after sex differentiation.

关 键 词：翘嘴鳜 NLRP3基因 分子标记 序列分析 表达分析 

分 类 号：Q78[生物学—分子生物学] S917.4[农业科学—水产科学]