尼罗罗非鱼
TNF-R1 和TNF-R2 基因克隆及表达Cloning and expression analysis of Nile tilapia
TNF-R1 andTNF-R2 gene- 中山大学学报(自然科学版) 2020年59卷第5期 页码：144-155
- 作者机构：
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/ 广东省重要经济鱼类健康养殖工程技术研究中心，广东 广州 510275
- 作者简介：
叶航宇（1995年生），男；研究方向：鱼类生理和鱼类免疫学；E-mail: yehy23@ mail2.sysu.edu.cn
吴金英（1963年生），女；研究方向：鱼类生理和鱼类免疫学；E-mail: lsswjy@mail.sysu.edu.cn
- 基金信息：现代农业产业技术体系专项资金(CARS-46);广州市科技计划重点项目(201904020043);现代农业人才支撑计划项目（2016-2020）
DOI：10.13471/j.cnki.acta.snus.2020.01.09.2020E002
中图分类号： Q17纸质出版日期：2020-09-25，
收稿日期：2020-01-09，
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叶航宇,葛岩岩,吴金英等.尼罗罗非鱼TNF-R1和TNF-R2基因克隆及表达[J].中山大学学报(自然科学版),2020,59(05):144-155.
YE Hangyu,GE Yanyan,WU Jinying,et al.Cloning and expression analysis of Nile tilapia TNF-R1 and TNF-R2 gene[J].Acta Scientiarum Naturalium Universitatis Sunyatseni,2020,59(05):144-155.
叶航宇,葛岩岩,吴金英等.尼罗罗非鱼TNF-R1和TNF-R2基因克隆及表达[J].中山大学学报(自然科学版),2020,59(05):144-155. DOI： 10.13471/j.cnki.acta.snus.2020.01.09.2020E002.
YE Hangyu,GE Yanyan,WU Jinying,et al.Cloning and expression analysis of Nile tilapia TNF-R1 and TNF-R2 gene[J].Acta Scientiarum Naturalium Universitatis Sunyatseni,2020,59(05):144-155. DOI： 10.13471/j.cnki.acta.snus.2020.01.09.2020E002.
摘要
为揭示TNF-α受体基因
TNF-R1
和
TNF-R2
在尼罗罗非鱼（
Oreochromis niloticus
）免疫过程的作用，本研究利用RACE技术获得了
TNF-R1
和
TNF-R2
的cDNA。结果表明，
TNF-R1
cDNA全长2 275 bp，ORF区1 335 bp，编码444个氨基酸；
TNF-R2
cDNA全长1 719 bp，ORF区1 476 bp，编码491个氨基酸。TNF-R1含3个富含半胱氨酸结构域（cysteine-rich domain
CRD）和一个死亡结构域（death domain
DD），而TNF-R2有4个CRD，缺少DD。系统进化分析表明：其
TNF-R1
和
TNF-R2
分别与其他脊椎动物
TNF-R1
和
TNF-R2
基因同源性很高，在进化树中
TNF-R1
、
TNF-R2
各自聚类且两者之间分支明显。实时荧光定量PCR结果表明：
TNF-R1
和
TNF-R2
在该鱼体的不同组织中均有表达，其中在鳃、头肾和脾脏中表达水平最高，而在肌肉、垂体中表达量低。用脂多糖（LPS）和多聚肌苷酸-胞苷酸（Poly Ⅰ:C）分别刺激离体孵育的该鱼头肾和脾脏白细胞可不同程度上调
TNF-α
、
TNF-R1
和
TNF-R2
的表达。无乳链球菌（
Streptococcus agalactiae
）感染该鱼可显著上调头肾和脾脏中3种基因的表达。本研究对揭示TNF-R1和TNF-R2免疫调控机制，提高罗非鱼健康养殖水平具有一定参考价值。
Abstract
To reveal the role of TNF-α receptor genes
TNF-R1
and
TNF-R2
in the Nile tilapis (
Oreochromis niloticus
) immune processes
we cloned the full-length cDNA sequences of
TNF-R1
and
TNF-R2
from Nile tilapia by RACE. Sequence analysis showed that the full length of
TNF-R1
cDNA was 2 275 bp
containing an open reading frame of 1 335 bp
and encoding a protein with 444 amino acids; the full length of
TNF-R2
cDNA was 1 719 bp
containing an open reading frame of 1 476 bp
and encoding a protein with 491 amino acids. Protein structure prediction showed that TNF-R1 have three cysteine-rich domains and an intracellular death domain
while TNF-R2 have four cysteine-rich domains and without death domain. Systematic evolutionary analysis showed that the tilapia
TNF-R1
and
TNF-R2
were highly homologous with those of other vertebrates. Phylogenetic tree showed that
TNF-R1
and
TNF-R2
were clustered separately. By qRT-PCR
the highest expression levels of
TNF-R1
and
TNF-R2
mRNA were found in the gill
head kidney and spleen
and the lowest levels were in pituitary and muscle. The expression of
TNF-α
TNF-R1
and
TNF-R2
genes can be increased to varying degrees by stimulating the tilapia kidneys and spleen white cells incubated with LPS and Poly Ⅰ:C
respectively. The expressions of three genes in the head
kidney and spleen can be increased after intra-peritoneal injection of
Streptococcus agalactiae
. This study has some reference value to show the immune regulation mechanisms of
TNF-R1
and
TNF-R2
to improve the level of healthy breeding of tilapia.
关键词
TNF-αTNF-R1TNF-R2罗非鱼免疫无乳链球菌
Keywords
TNF-αTNF-R1TNF-R2Oreochromis niloticusimmunityStreptococcus agalactiae
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