南方农业学报2025，Vol.56Issue(1)：305-313,9.DOI:10.3969/j.issn.2095-1191.2025.01.027
白斑狗鱼LADD 2基因克隆及其组织表达特征分析
Cloning and tissue expression characteristics analysis of LADD 2 gene in Esox lucius
摘要
Abstract
[Objective]To clone LADD 2 gene of Esox lucius and analyze its tissue expression characteristics,which could provide theoretical basis for revealing the role of LADD 2 gene in the gonadal development of E.lucius.[Method]Using the gonadal transcriptome data of E.lucius,the highly expressed gene LADD 2 in 126-day-old female E.lucius was screened.The coding region(CDS)sequence of LADD 2 gene was cloned,and bioinformatics analysis was performed using online tools such as DNAMAN 9.0,NCBI ORF finder,BLAST,ClustalX,ExPASy,ProtParam,PSORT II,SOMPA and SWISS-MODEL.Based on the amino acid sequences of fish ladder-type lectin homologs and C-type lectin re-ceptors,a phylogenetic tree was constructed using the neighbor-joining(NJ)method in MEGA 5.1.The relative expres-sion levels of LADD 2 gene in the gonads,intestines,muscles,liver,gills,brain,kidneys and head kidneys of E.lucius at 126,188,and 320 day-old were detected using real-time fluorescence quantitative PCR.[Result]The CDS sequence of LADD 2 gene in E.lucius was successfully cloned,with a total length of 609 bp,encoding 85 amino acids residues.The protein molecular weight was 9.78 kD,the theoretical isoelectric point(pI)was 7.30,and the molecular formula was C8635H13504N2334O2404S115,indicating a hydrophilic protein.The LADD 2 protein had the highest content of leucine(11.86% ),and the lowest content of tryptophan(2.28% ).Phosphorylation site prediction showed that LADD 2 protein contained 147 serine sites,98 threonine sites and 44 tyrosine sites.Protein structure prediction revealed that LADD 2 pro-tein mainly consisted of random coils β-turns,α-helices and β-folds,with 2 conserved PRT1 domains.Amino acid se-quence homology analysis showed that the similarity between the LADD 2 protein of E.lucius and that of Dallia pectora-lis was the highest at 99% .The phylogenetic tree indicated that LADD 2 protein of E.lucius clustered with fish ladder-type lectin homologs and was closely related to Salmo salar,Oncorhynchus kisutch and Coregonus clupeaformis.Real-time fluorescence quantitative PCR results showed that the relative expression of LADD 2 gene in the ovaries of E.lucius gradually decreased with developmental time,and the difference in relative expression between the ovaries and testes be-came smaller.Additionally,LADD 2 gene was expressed in the gills and brain.[Conclusion]The function of LADD 2 gene exhibits certain conservation and stability during evolution.LADD 2 protein may play an important role in the early ovarian development of E.lucius and may have immune activity in gills and brain during later developmental stages.LADD 2 protein may possess immune functions,thereby protecting the hepatopancreatic development of E.lucius.关键词
白斑狗鱼/LADD 2基因/梯型凝集素/组织表达特征Key words
Esox lucius/LADD 2 gene/ladder-type lectin/tissue expression characteristics分类
农业科技引用本文复制引用
刘祎,王顺哲,刘璎慧,杨倩,汪泳昌,孙浩然,李胜忠,张俊杰..白斑狗鱼LADD 2基因克隆及其组织表达特征分析[J].南方农业学报,2025,56(1):305-313,9.基金项目
国家自然科学基金项目(32260915) （32260915）
新疆自然科学基金项目(2022D01A66) National Natural Science Foundation of China(32260915) （2022D01A66）
Xinjiang Natural Science Foundation(2022D01A66) （2022D01A66）
