中国畜牧兽医2023，Vol.50Issue(12)：4783-4792,10.DOI:10.16431/j.cnki.1671-7236.2023.12.001
绵羊SMAD4基因克隆、生物信息学分析及组织表达研究
Cloning,Bioinformatics Analysis and Tissue Expression of SMAD4 Gene in Sheep
摘要
Abstract
[Objective]This study was aimed to clone SMAD homolog 4(SMAD4)gene and explore its biological function in sheep,so as to provide a basis for studying the regulatory role of SMAD4 gene in the growth and development of hair follicles.[Method]Using the cDNA of skin in Small-tailed Han sheep as a template,the CDS region of SMAD4 gene was amplified by PCR and cloned,and was sequenced for bioinformatics analysis.Real-time quantitative PCR was used to detect the expression of SMAD4 gene in heart,liver,spleen,kidney,skin and muscle in Small-tailed Han sheep and Xinji Fine wool sheep.[Result]The CDS region of SMAD4 gene in Small-tailed Han sheep was successfully cloned,which contained 1 662 bp and encoded 553 amino acids.Phylogenetic tree analysis showed that the nucleotide sequence of SMAD4 gene in sheep was the closest to Capra hircus and Bos mutus,and the most distant with Gallus gallus.Bioinformatics analysis found that SMAD4 protein was a non-transmembrane protein without signal peptides,with phosphorylation and glycosylation sites,and belonged to hydrophilic proteins,mainly in nucleus.The secondary structure of SMAD4 protein was dominated by random coil,which was consistent with the tertiary structure.The results of protein interaction analysis showed that SMAD4 protein interacted with SMAD2,SMAD1,TGFBR1,TGFBR2,SMAD3 and other proteins.Real-time quantitative PCR results showed that SMAD4 gene was widely expressed in sheep.Compared with Xinji Fine wool sheep,the expression of SMAD4 gene in heart,spleen and skin of Small-tailed Han sheep were significantly increased(P＜0.05),but the expression of SMAD4 gene in lung and muscle were significantly decreased(P＜0.05).[Conclusion]The CDS region of SMAD4 gene in sheep was successfully cloned,which was expressed in various tissues of sheep,and there were significant differences in heart,spleen,skin,lung and muscle between Small-tailed Han sheep and Xinji Fine wool sheep.The results provided a theoretical basis for further exploration of the regulation of hair follicle development by SMAD4 gene in sheep.关键词
绵羊/SMAD4基因/克隆/生物信息学/表达Key words
sheep/SMAD4 gene/clone/bioinformatics/expression分类
畜牧业引用本文复制引用
王丹,张新玉,付佳棋,贾琪,张双双,崔成都,张立春,孙福亮..绵羊SMAD4基因克隆、生物信息学分析及组织表达研究[J].中国畜牧兽医,2023,50(12):4783-4792,10.基金项目
国家自然科学基金项目(32060781) （32060781）
吉林省教育厅项目(JJKH20220543KJ) （JJKH20220543KJ）
