摘要
Galectin-8是半乳糖凝集素家族的成员,对β-半乳糖苷类能够特异性识别,具有多种生物学功能。为研究杉虎斑(Epinephelus fuscoguttatus♀×E. polyphekadion♂)Galectin-8基因的序列特征、组织表达情况与饲料中添加阿魏酸(ferulic acid,FA)对其表达的影响,使用cDNA末端快速扩增技术(rapid amplification of cDNA ends,RACE)首次克隆得到了其全长cDNA序列。该序列长1 368 bp,其中5’UTR为16 bp,3’UTR为394 bp,开放阅读框(ORF)长960 bp,共编码319个氨基酸。该氨基酸序列在N端和C端上各具有一个糖识别结构域(carbohydrate binding domain,CRD),分别由136和133个氨基酸组成,由长为36个氨基酸的连接肽所连接,并包含两个保守基序H-NPR和WG-EE。同源性与系统发育进化树分析表明,杉虎斑Galectin-8与鞍带石斑鱼的一致性最高,为98.43%,并与其他硬骨鱼类聚为一支。RT-qPCR结果显示,Galectin-8在杉虎斑脾脏中表达量最高,其次是中肾、心脏、肠道、肝脏、头肾和鳃,而在脑、胃、皮肤和肌肉组织中微量表达。饲料中添加40-320 mg/kg的FA时,肠道、肝脏、头肾和脾脏的Galectin-8表达量显著上调(P <0.05),这表明FA可作为免疫刺激剂增强杉虎斑的先天性免疫。然而,杉虎斑Galectin-8的免疫功能尚不清,需要进一步的研究来阐明,以了解其在抵抗病原体入侵中的作用。
Galectin-8 is a member of the galectin family,and binds with β-galactoside by specific recognition with many biological functions. The present study was conducted to investigate Galectin-8 sequence characterization,its tissue distribution,and the effect of dietary ferulic acid(FA)supplementation on the Galectin-8 gene expression in hybrid grouper(Epinephelus fuscoguttatus ♀ × E. polyphekadion ♂). The technology of rapid amplification of cDNA ends(RACE)was utilized to obtain the full length(1 368 bp)of hybrid grouper Galectin-8 cDNA for the first time. The cDNA was composed of 5’ untranslated region(5’ UTR)(16 bp),3’ UTR(394 bp),and open reading frame(ORF)of 960 bp that encoded 319 amino acids. The amino acid sequence of galectin-8 possessed two carbohydrate recognition domains(CRDs)at N-and C-terminus composed of 136 and 133 amino acids,respectively,which were connected by a linker peptide of 36 amino acids and contained two conserved H-NPR and WG-EE motifs. Homology and phylogenetic tree analysis indicated that hybrid grouper Galectin-8 demonstrated the highest identity(98.43%)with E. fuscoguttatus,and was clustered with other teleosts. Based on RT-qPCR,tissue distribution analysis revealed that Galectin-8 showed the highest expressions in the spleen,followed by mid kidney,heart,intestine,liver,head kidney,and gill,yet the trace expression in the brain,stomach,skin,and muscle. Moreover,dietary FA supplementation at level of 40-320 mg/kg considerably up-regulated the Galectin-8 transcriptional level of the intestine,liver,head kidney,and spleen in hybrid grouper(P < 0.05),which suggested that FA served as an immunostimulant to improve its innate immunity. Nevertheless,Galectin-8 immune functions of the hybrid grouper are unclear,and therefore further studies are needed to illustrate its role in defense against pathogen invasion.
作者
付伟杰
邝杰华
罗君
黄建盛
陈有铭
陈刚
FU Wei-jie;KUANG Jie-hua;LUO Jun;HUANG Jian-sheng;CHEN You-ming;CHEN Gang(College of Fisheries,Guangdong Ocean University,Zhanjiang 524088;Guangdong Blue Granary Seed Industry Co.,Ltd.,Zhanjiang 524000)
出处
《生物技术通报》
CAS
CSCD
北大核心
2022年第12期312-323,共12页
Biotechnology Bulletin
基金
国家重点研发计划“蓝色粮仓”重点专项(2020YFD0900200)
广东省科技重点研发计划(2016B0201009)。
关键词
杉虎斑
Galectin-8
阿魏酸
基因克隆
表达分析
Epinephelus fuscoguttatus♀×E.polyphekadion♂
Galectin-8
ferulic acid
gene cloning
expression analysis
