七彩神仙鱼BTN1A1基因克隆及其在嗜水气单胞菌胁迫下的表达分析
Cloning of BTN1A1 Genes from Symphysodon aequifasciatus and Analysis of Its Expression under Aeromonas hydrophila Stress
何恺轩 1温彬 1高建忠 1陈再忠1
作者信息
- 1. 上海海洋大学水产与生命学院/农业农村部淡水水产种质资源重点实验室/水产种质资源发掘与利用教育部重点实验室/水产科学国家级实验教学示范中心,上海 201306
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摘要
[目的]克隆七彩神仙鱼(Symphysodon aequifasciatus)嗜乳脂蛋白1A1(Butyrophilin 1A1,BTN1A1)基因,分析其在病原刺激下的表达模式,为了解七彩神仙鱼BTN1A1基因功能提供依据.[方法]利用生物信息学对七彩神仙鱼2个BTN1A1基因(BTN1A1-1和BTNA41-2)进行结构和进化分析.根据前期获得的七彩神仙鱼皮肤转录组数据,选取BTN1A1-1和BTN1A1-2基因的CDS区设计引物进行克隆.采用qRT-PCR分析BTN1A1在各组织中的表达及嗜水气单胞菌刺激下的表达模式.[结果]BTN1A1-1和BTN1A1-2的ORF序列长度为894、1 275 bp,分别编码298、424个氨基酸.BTN1A1-1和BTN1A1-2蛋白均具有1个信号肽、1个Ig结构域、1个lg_like结构域和1个跨膜结构域的经典结构,BTN1A1-2还具有1个胞质结构域.七彩神仙鱼BTN1A1-1与慈鲷科其他鱼类相似性较高,与尼加拉瓜湖始丽鱼(Archocentrus centrarchus)BTNIAI对应氨基酸序列同源性最高、为94.14％.但BTN1A1-2与其他鱼类发生分离,形成独特分支.BTN1A1-1和BTN1A1-2在七彩神仙鱼各组织中均有表达,但在鳃、肠道、皮肤等与免疫相关的组织中表达量相对较高.七彩神仙鱼BTN1A1-1在嗜水气单胞菌胁迫后表达量先显著下降后上升,而BTN1A1-2表达量则先显著上升后下降.[结论]七彩神仙鱼BTN1A1-1基因相对保守,而BTN1A1-2基因在进化上较为特殊.二者在免疫刺激下的差异表达暗示其可能在七彩神仙鱼免疫防御中发生功能分化.
Abstract
[Objective]Butyrophilin 1A1(BTN1A1)gene from Symphysodon aequifasciatus was cloned and its express pattern under pathological infection was analyzed,to provide basis for better understanding of the function of BTN1A1 gene in S.aequifasciatu.[Method]The structure and evolutionary characteristics of two BTN1A1 genes(BTN1A1-1 and BTN1A1-2)were analyzed by bioinformatics analysis.Based on the transcriptome data of the skin of the S.aequifasciatu obtained at early stage,the CDS regions of BTN1A1-1 and BTN1A1-2 genes were used to design the primers for gene cloning.The expressions of BTN1A1-1 gene in various tissues and its expression pattern under the stimulus of Aeromonas hydrophila were analyzed by qRT-PCR.[Result]The ORF regions of BTN1A1-1 and BTN1A1-2 were 894 and 1 275 bp,encoding 298 and 424 amino acids,respectively.Both BTN1A1-1 and BTN1A1-2 had a classical structure of BTN,which contained a signal peptide,an Ig domain,an lg_like domain and a transmembrane domain.BTN1A1-2 also had a cytoplasmic domain.BTN1A1-1 showed a closer relationship with the genes found in cichlid fish.It shared the highest homology of 94.14％with the corresponding amino acid sequences of Archocentrus centrarchus;while BTN1A1-2 showed lower similarity with other types of fish,forming a special branch.The expressions of BTN1A1-1 and BTN1A1-2 were highly in gill,intestine and skin of S.aequifasciatu.The expression of BTN1A1-1 decreased significantly and then increased after the infection of A.hydrophila,while the expression of BTN1A1-2 increased significantly and then decreased.[Conclusion]The BTN1A1-1 gene was more evolutionarily conservative than BTN1A1-2.The differential expressions of BTN1A1-1 and BTN1A1-2 in response to immune stimulation suggests that they may be functionally differentiated in the immune defence of S.aequifasciatus.
关键词
七彩神仙鱼/嗜乳脂蛋白/嗜水气单胞菌/免疫调节/基因表达Key words
Symphysodon aequifasciatus/butyrophilin/Aeromonas hvdrophila/immunomodulation/gene expression引用本文复制引用
基金项目
上海市自然科学基金(20ZR1423600)
上海海洋大学科技发展专项(A2-2006-22-2002)
国家自然科学基金(31902376)
出版年
2024
NETL
NSTL
万方数据