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2025年4月4日 星期五
农业生物技术学报  2021, Vol. 29 Issue (4): 732-740    DOI: 10.3969/j.issn.1674-7968.2021.04.011
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
牙鲆NTF3基因克隆与表达分析
陈志鹏1,2, 朱颖2, 王磊2, 胡国斌3, 陈松林2,*
1 中国海洋大学 水产学院,青岛 266071;
2 中国水产科学研究院 黄海水产研究所/青岛海洋科学与技术试点国家实验室海洋渔业科学与食物产出过程功能实验室, 青岛 266071;
3 中国海洋大学 海洋生命学院,青岛 266071
Cloning and Expression Analysis of NTF3 Gene in Flounder (Paralichthys olivaceus)
CHEN Zhi-Peng1,2, ZHU Ying2, WANG Lei2, HU Guo-Bin3, CHEN Song-Lin2,*
1 Fisheries College, Ocean University of China, Qingdao 266071, China;
2 Yellow Sea Fisheries Research Institute, Qingdao National Laboratory of Marine Science and Technology Pilot Laboratory of Marine Fishery Science and Food Production Process Functional Laboratory, Chinese academy of fishery sciences, Qingdao 266071, China;
3 College of Marine Life, Ocean University of China, Qingdao 266071, China
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摘要 造成牙鲆(Paralichthys olivaceus)工厂化养殖中大规模死亡的主要病原之一是迟缓爱德华氏菌(Edwardsiella tarda),其有感染力极强、致死率极高的特点,严重影响牙鲆水产养殖产业的健康发展。神经营养因子3 (neurotrophin3, NTF3)是一种神经营养因子,可能参与免疫系统的调节。为研究NTF3在牙鲆先天免疫中的功能,本研究克隆了牙鲆NTF3基因的全长cDNA序列,其长度为1 411 bp,其中,5'非编码区长89 bp,3'非编码区长428 bp,ORF长894 bp。基因ORF编码297个氨基酸。预测分子量(molecular weight, Mw)约为33.73 ku,理论等电点为9.56。同源比对发现,NTF3基因在不同物种间的相似度较高,牙鲆与高体鰤(Seriola dumerili) NTF3基因的同源性最高,为90.91%。NTF3基因在牙鲆健康组织(血液, 肝脏, 脾脏, 肾脏, 肠, 皮肤, 鳃, 心脏和脑)中均有表达,皮肤中的表达量最高,血液中次之。为进一步探究牙鲆感染迟缓爱德华氏菌过程中NTF3相对表达量的变化,本研究对健康牙鲆进行了迟缓爱德华氏菌感染实验,比较了NTF3在4种组织(脾脏, 肝脏, 肠, 肾脏)不同感染时间点的相对表达量,在脾脏、肝脏、肠中,牙鲆感染迟缓爱德华氏菌后NTF3基因的表达量均出现上调,但在肾脏中,NTF3的表达量呈现下调趋势。上述结果表明,NTF3基因参与了牙鲆抵抗迟缓爱德华氏菌感染的免疫过程,而病菌感染后不同组织中 NTF3基因的相对表达量差异的结果为今后更深入研究NTF3基因在牙鲆免疫应答过程中发挥的作用提供了基础研究数据。
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陈志鹏
朱颖
王磊
胡国斌
陈松林
关键词 牙鲆迟缓爱德华氏菌神经营养因子3 (NTF3)基因表达    
AbstractEdwardsiella tarda is the main pathogen causing large-scale death of flounder (Paralichthys olivaceus) in factory farming. Its infectivity is extremely strong. The mortality rate of infected flounder with E. tarda is very high, which seriously impedes the development of flounder industry. In order to study the function of neurotrophin 3 (NTF3) in the innate immunity of P. olivaceus, the full-length cDNA sequence of P. olivaceus NTF3 gene was cloned, with the total length 1 411 bp, in which the 5' noncoding region is 89 bp and the 3' non-coding region is 428 bp.The open reading frame (ORF) of this gene was 894 bp.Which encoded 297 amino acids. The predicted molecular weight (Mw) was about 33.73 ku, and the theoretical isoelectric point (pI) was 9.56. The homology comparison found that the NTF3 gene had a high similarity with different species, and the highest homology was 90.91% with the NTF3 gene of amberjack (Seriola dumerili). The NTF3 gene was expressed in lots of healthy tissues of flounder (blood, liver, spleen, kidney, intestine, skin, gill, heart and brain). The highest expression level was tested in skin followed by blood. To further study the relative expression of NTF3 during E. tarda infection, a bacterial infection experiment was conducted on healthy flounder, and then the relative expression of NTF3 in 4 different tissues (intestine, kidney, spleen and liver) and different time points were compared. It was found that, in the intestine, spleen and liver, the expression levels of NTF3 gene were up-regulated after bacterial stimulation, but in the kidney, the expression levels showed a downward trend. The above results indicated that the NTF3 gene may be involved in the immune process of P. olivaceus against the E.tarda infection, and the relative expression of NTF3 gene is different in different tissues infected by the bacteria, which provides basic research data for further study of the role of NTF3 gene in the immune response process of flounder.
Key wordsParalichthys olivaceus    Edwardsiella tarda    Neurotrophin 3 (NTF3)    Gene expression
收稿日期: 2020-07-14      出版日期: 2021-04-01
ZTFLH:  S917.4  
基金资助:山东省自然科学基金(ZR2016QZ003); 山东省泰山学者攀登计划专家项目; 中国水产科学研究院科技创新团队项目(2020TD20)
通讯作者: * chensl@ysfri.ac.cn   
引用本文:   
陈志鹏, 朱颖, 王磊, 胡国斌, 陈松林. 牙鲆NTF3基因克隆与表达分析[J]. 农业生物技术学报, 2021, 29(4): 732-740.
CHEN Zhi-Peng, ZHU Ying, WANG Lei, HU Guo-Bin, CHEN Song-Lin. Cloning and Expression Analysis of NTF3 Gene in Flounder (Paralichthys olivaceus). 农业生物技术学报, 2021, 29(4): 732-740.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2021.04.011     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2021/V29/I4/732
 
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