半滑舌鳎突触体相关蛋白29基因(SNAP29)克隆及其组织分布和时序表达分析OA北大核心CSTPCD
Cloning,Tissue Distribution and Temporal Expression Analysis of Synaptosome-associated Protein 29 Gene(SNAP29)in Cynoglossus semilaevis
为了分析半滑舌鳎突触体相关蛋白29 基因(SNAP29)的分子生物学特征、组织分布特点以及感染海藻希瓦氏菌后的时序表达规律,通过RT-PCR、生物信息学网站、qRT-PCR技术对半滑舌鳎SNAP29 基因进行了扩增、生信分析和表达特征研究.结果表明,半滑舌鳎SNAP29 基因CDS区长度为789 bp,编码 262 个氨基酸,理论等电点(pI)为5.39,分子质量为29.71781 ku,有2 个典型的SNARE结构域;二级结构主要由α-螺旋和无规卷曲组成,二级结构和三级结构基本一致;多序列比对结果显示,半滑舌鳎与欧洲鲈鱼SNAP29 基因相似性最高;SNAP29 基因在健康半滑舌鳎所测的各组织中均有表达,在鳃组织中表达最高,在脑组织中表达量最低;利用海藻希瓦氏菌人工感染半滑舌鳎后,SNAP29 基因在肠道组织、心脏组织和脑组织中表达量主要呈现上调趋势,在肝脏、头肾、脾脏组织中主要呈现下调表达,SNAP29 基因在健康半滑舌鳎组织中均有表达;海藻希瓦氏菌感染半滑舌鳎的肠道、脑、心脏、肝脏、头肾、脾脏组织中SNAP29 基因呈现差异表达.综上,SNAP29 可能参与机体免疫应答过程或病理生理过程.
The aim of this study was to analyze synaptosome associated protein 29 gene(SNAP29)in Cynoglos-sus semilaevis.The molecular biological characteristics,tissue distribution characteristics of SNAP29 gene and the temporal expression pattern after infection with Shewanella algae.The SNAP29 gene of C.semilaevis was amplified by RT-PCR,bioinformatics website and qRT-PCR,and its bioinformatics analysis and expression characteristics were studied.The results showed that the CDS region of SNAP29 gene was 789 bp in length,encoding 262 amino acids.The theoretical isoelectric point(pI)was 5.39,and the molecular weight was29.71781 ku.There were two typical SNARE domains.The secondary structure was mainly composed of α-helix and random coil,and the secondary structure and tertiary structure were basically the same.The results of multiple sequence alignment showed that the SNAP29 gene of C.semilaevis had the highest similarity with that of European perch.The SNAP29 gene was ex-pressed in all tissues of healthy C.semilaevis,with the highest expression in gill tissue and the lowest expression in brain tissue.After artificial infection of C.semilaevis with S.algae,the expression of SNAP29 gene was mainly up-regulated in intestinal tissue,heart tissue and brain tissue,and down-regulated in liver,head kidney and spleen tis-sue.SNAP29 gene was expressed in heart,brain,liver,intestinal tract,spleen,gill,middle kidney and head kidney of healthy C.semilaevis.The SNAP29 gene was differentially expressed in the intestine,brain,heart,liver,head kidney and spleen of C.semilaevis infected with S.algae.The above results indicate that SNAP29 may be involved in the body's immune response process or pathophysiological process.
吴茵茵;温思怡;韩卓然;孙敬锋
天津农学院,农业农村部智慧养殖重点实验室(部省共建),天津 300384||天津市水产生态及养殖重点实验室,天津农学院 水产学院 ,天津 300384天津市水产生态及养殖重点实验室,天津农学院 水产学院 ,天津 300384
水产学
半滑舌鳎SNAP29基因克隆qRT-PCR海藻希瓦氏菌
Cynoglossus semilaevisSNAP29Genetic cloningqRT-PCRShewanella algae
《华北农学报》 2024 (002)
231-238 / 8
国家自然科学基金项目(31972840);天津市海水养殖产业技术体系项目(ITTMRS2021007);农业农村部智慧养殖重点实验室(部省共建)开放基金项目(2023-TJAULSBF-2105)
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