关键词: 白来航鸡; Galectin-1基因; 基因克隆; 生物信息学; 组织表达

Abstract: 【Objective】 This study was aimed to clone the galectin-1 (Gal-1) gene in chickens,analyze its encoded protein by bioinformatics techniques,and detect its expression in different tissues in White Leghorn chickens,in order to provide scientific basis for further elucidating its in antiviral effect.【Method】 PCR was used to amplify the complete CDS region of Gal-1 gene with the spleen cDNA as template,and the similarity alignment and phylogenetic tree construction were carried out.Bioinformatics softwares were used to predict the physicochemical properties,hydrophilicity/hydrophobicity,transmembrane domain,signal peptide,modified structure,conserved domains and advanced structures of Gal-1 protein.Real-time quantitative PCR was used to detect the expression of Gal-1 gene in heart,liver,spleen,lung,kidney,brain,glandular stomach,muscular stomach,duodenum,jejunum,cecum,rectum,pectorales and leg muscles tissnes.【Result】 The length of CDS region of chicken Gal-1 gene was 408 bp,encoding 135 amino acids.The similarity alignment results showed that the similarity of Gal-1 gene nucleotide sequence of White Leghorn chickens with Meleagris gallopavo,Anas platyrhynchos and Taeniopygia guttata were 97.1%,88.4% and 82.2%,respectively.The phylogenetic tree analysis showed that White Leghorn chickens had the closest relationship with Meleagris gallopavo.The relative molecular weight of Gal-1 protein was 15.06 ku,the isoelectric point was 6.57,the instability coefficient was 36.05 and the lipid coefficient was 74.30,and the average hydrophilic index was ―0.259.Gal-1 protein had no signal peptide and no transmembrane domain,while had two obvious hydrophilic domains,and the coding protein was stable and hydrophilic.The prediction of secondary structure showed that the Gal-1 protein was dominated by random coil (45.93%) and extended chain (41.48%),and the prediction of tertiary structure was consistent with that of secondary structure.Real-time quantitative PCR results showed that Gal-1 gene mRNA was widely expressed in tissues of White Leghorn chickens,with the highest expression level in the lung and the lowest in the brain.【Conclusion】 In this study,the CDS region sequence of Gal-1 gene was successfully cloned,and it was widely expressed in 14 tissues such as heart and liver of White Leghorn chickens,which could provide references for further study on the function of Gal-1 protein in chickens.

Key words: White Leghorn chickens; Galectin-1 gene; gene cloning; bioinformatics; tissue expression