合作猪CXCL12基因克隆、生物信息学分析及组织表达研究

doi:10.16431/j.cnki.1671-7236.2025.06.003

摘要/Abstract

摘要： 【目的】对合作猪基质细胞衍生因子-1(stromal cell-derived factor 1，SDF-1(CXCL12))基因进行克隆及生物信息学分析，并检测其在不同组织中的表达水平，为研究CXCL12基因的功能奠定基础。【方法】以合作猪心脏组织cDNA为模板，采用PCR扩增、克隆CXCL12基因CDS区序列并测序，对所获序列进行相似性比对及系统进化树构建。利用生物信息学软件对合作猪CXCL12蛋白进行理化性质及结构功能分析。运用实时荧光定量PCR检测CXCL12基因在合作猪各组织内的表达水平。【结果】合作猪CXCL12基因CDS区全长351 bp，编码116个氨基酸；与马的亲缘关系最近，与羚羊的亲缘关系最远。CXCL12蛋白不存在跨膜区和信号肽，但存在1个低复杂度的SCY结构域，亲水性指数为―0.347，属于亲水性蛋白，包含12个磷酸化位点；二级结构主要为无规则卷曲(54.31%)和α-螺旋(32.76%)，三级结构与二级结构相一致；该蛋白主要分布于细胞核和线粒体，并与CCL21、CXCL9等蛋白存在相互作用。实时荧光定量PCR结果显示，CXCL12基因在合作猪肝脏中的表达量极显著高于其他组织(P＜0.01)，在肾脏和心脏中的表达量极显著低于其他组织(P＜0.01)。【结论】本研究成功克隆合作猪CXCL12基因CDS区，编码116个氨基酸，主要分布在细胞核和线粒体中，是一种稳定、亲水且无跨膜结构的分泌蛋白。CXCL12基因在合作猪肝脏中的表达量最高。本研究结果可为进一步揭示CXCL12基因调控合作猪免疫机制提供参考。

关键词: 合作猪; CXCL12基因; 克隆; 生物信息学; 表达

Abstract: 【Objective】 This study was aimed to clone and perform bioinformatics analysis on the stromal cell-derived factor-1 (SDF-1/CXCL12) gene in Hezuo pigs，and detect the expression of CXCL12 gene in different tissues,establishing a foundation for further functional investigation of CXCL12 gene.【Method】 Using cDNA from the heart of Hezuo pigs as a template,the CXCL12 gene CDS sequence was amplified via PCR,cloned and sequenced,the similarity alignment and phylogenetic tree construction for the obtained sequence were performed.Bioinformatics software was used to analyze the physicochemical property and structural function of CXCL12 protein.The expression of CXCL12 gene in different tissues of Hezuo pigs was detected by Real-time quantitative PCR.【Result】 The full-length CDS sequence of CXCL12 gene in Hezuo pigs was 351 bp,encoding 116 amino acids.CXCL12 gene in Hezuo pigs was most closely related to Equus cabllus,and was distantly related to Oryx dammah.CXCL12 protein lacked transmembrane regions and signal peptide,included a low complexity region SCY,with a hydrophilicity index of ―0.347,it belonged to hydrophilic proteins.CXCL12 protein contained 12 phosphorylation sites.The secondary structure was predominantly compose of random coil (54.31%) and alpha helix (32.76%),which was consistent with the tertiary structure.The protein was primarily located in nucleus and mitochondria,and interacted with proteins such as CCL21 and CXCL9.Real-time quantitative PCR results showed that the expression of CXCL12 gene in liver of Hezuo pigs was extremely significantly higher than that in other tissues (P＜0.01),and the expression in kidney and heart was extremely significantly lower than that in other tissues (P＜0.01).【Conclusion】 This study successfully cloned the CDS sequence of CXCL12 gene in Hezuo pigs,encoding 116 amino acids,mainly distributed in nucleus and mitochondria.It was a stable,hydrophilic and secretory protein without transmembrane structures.The highest expression of CXCL12 gene was observed in liver of Hezuo pigs.The results provided a reference for further exploring the regulatory roles of CXCL12 gene in immune mechanisms of Hezuo pigs.

Key words: Hezuo pigs; CXCL12 gene; cloning; bioinformatics; expression

中图分类号:

S813.3
Q78

杨泉, 李晓, 闫尊强, 王鹏飞, 黄晓宇, 高小莉, 杨巧丽, 滚双宝, 杨姣姣. 合作猪CXCL12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, 2025, 52(6): 2482-2493.

YANG Quan, LI Xiao, YAN Zunqiang, WANG Pengfei, HUANG Xiaoyu, GAO Xiaoli, YANG Qiaoli, GUN Shuangbao, YANG Jiaojiao. Cloning,Bioinformatics Analysis and Tissue Expression of CXCL12 Gene in Hezuo Pigs[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(6): 2482-2493.

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