关中奶山羊PURB基因克隆、生物信息学分析及组织表达研究OA

This study aimed to clone the coding sequence(CDS)of the purine-rich element-binding protein B(PURB)gene in Guanzhong dairy goats,perform bioinformatic characterization,determine its tissue expression profile,and preliminarily investigate its potential role in mammary gland development.Using cDNA synthesized from mammary gland tissue of Guanzhong dairy goats as the template,the CDS of the PURB gene was amplified by PCR and sequenced.Sequence characteristics were analyzed using bioinformatic methods,and the expression levels of PURB in the mammary gland,uterus,heart,kidney,spleen,and liver were determined by quantitative real-time PCR.The results showed that the full-length CDS of the PURB gene was successfully cloned.It was 915 bp in length and encoded a protein of 304 ami-no acids.Amino acid sequence alignment demonstrated that the PURB protein is highly conserved among different species.Phylogenetic analysis indicated that the goat PURB protein is more closely related to those of sheep and cattle than to those of horses and dromedary camels.The predicted PURB protein has a molecular formula of C1426 H2228 N420 O454 S5 and was identified as a hydrophilic,non-transmembrane protein lacking a signal peptide.It contains 21 potential phosphorylation sites and is predominantly localized in the nucleus.Secondary structure prediction showed that the PURB protein is mainly composed of β-sheets,α-helices,and random coils,and the predicted tertiary structure was consistent with the secondary structure.The PURB gene exhibited tissue-specific expression,with significantly higher expression levels in the ute-rus and spleen than in the other tissues examined(P＜0.05),whereas it was scarcely expressed in the heart.In conclusion,the CDS of the PURB gene was successfully cloned from Guanzhong dairy goats,and its bioinformatic characteristics and tissue expression profile were characterized,providing a reference for further functional studies of the PURB gene in this breed.