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梅花鹿DLX5基因克隆及表达分析

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为研究梅花鹿(Cervus nippon)DLX5基因的结构和功能,进一步探究其与梅花鹿茸角骨化机制间的关系,采用RT-PCR技术对梅花鹿DLX5基因进行克隆,获得包含全部编码区的cDNA序列,对该基因的氨基酸序列进行生物信息学分析并构建系统进化树,通过KEGG富集分析其信号通路,运用实时荧光定量RT-PCR检测该基因在鹿茸生长不同时期的表达情况.结果表明:梅花鹿DLX5基因编码区长为870 bp,共编码289个氨基酸.DLX5蛋白为可溶性的不稳定蛋白,有2个保守结构域,主要定位于细胞核.梅花鹿DLX5蛋白与许多不同物种来源的DLX5蛋白氨基酸序列有较高相似度,比较保守.DLX5蛋白二级结构中无规则卷曲占比最大(78.89%),其后依次是α-螺旋、延伸链,占比分别为16.96%和4.15%.DLX5基因主要的作用通路为TGFβ信号通路、MAPK信号通路和Wnt信号通路等.实时荧光定量RT-PCR结果表明,DLX5基因在鹿茸生长后期(三杈茸)表达量显著增高,这种上调表达暗示了其在鹿茸骨化过程中发挥重要作用,说明其可能是鹿茸骨化相关候选基因.
Cloning and Expression Analysis of DLX5 Gene in Sika Deer
In order to study the structure and function of DLX5 gene in sika deer(Cervus nippon),and to further explore the underlying ossification mechanism of sika antler velvet,DLX5 gene of sika deer was cloned by RT-PCR.The cDNA se-quence containing all coding regions was obtained.The amino acid sequence of the gene was analyzed by using bioinfor-matics method and the phylogenetic tree was constructed.The signaling pathway was analyzed by KEGG enrichment,and finally real-time RT-PCR was used to detect the expression of DLX5 gene in different growth periods of deer antler velvet.The results showed that the coding region of DLX5 gene in sika deer was 870 bp,and a total of 289 amino acids were en-coded.DLX5 protein was a soluble,unstable protein with two conserved domains and was mainly localized to the nucleus.Sika deer DLX5 protein had a high similarity with the amino acid sequence of DLX5 proteins derived from the different spe-cies,which was relatively conservative.Irregular curl accounted for the largest proportion in the secondary structure of DLX5 protein,and followed by α-helix and extended chain.Irregular curl,α-helix and extended chain was 78.89%,16.96%and 4.15%,respectively.The main pathways of DLX5 gene were TGFβ signaling pathway,MAPK signaling path-way and Wnt signaling pathway.The real-time RT-PCR results showed that the expression level of the gene was significantly increased in the late stage of deer antler growth(trident velvet),and the upregulated expression suggested that it played an important role in the process of deer antler ossification,which may be a candidate gene for deer antler ossification.

Sika deer(Cervus nippon)DLX5 geneGene cloningBioinformatics analysisReal-time RT-PCR

王鹏、刘方政、李萱博、王春花、于海浩、夏彦玲

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东北林业大学野生动物与自然保护地学院,哈尔滨,150040

黑龙江工程学院计算机科学与技术学院,哈尔滨,150050

梅花鹿 DLX5基因 基因克隆 生物信息学分析 实时荧光定量RT-PCR

国家级大学生创新训练项目黑龙江省哲学社会科学研究规划项目

20221022517021TQB117

2024

野生动物学报
东北林业大学 中国动物园协会

野生动物学报

北大核心
影响因子:0.271
ISSN:1000-0127
年,卷(期):2024.45(1)
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