塔里木马鹿WNT5B基因序列特征及其在鹿茸组织中的表达特征  

Sequence Characterization of WNT5B Gene and Its Expression Profile in Antler Tissues of Tarim Red Deer

作　　者：向欣 杨晨晨 林川 陈红 吕皓[1,2] 韩春梅 XIANG Xin;YANG Chenchen;LIN Chuan;CHEN Hong;LU Hao;HAN Chunmei(College of Animal Science and Technology,Tarim University,Aral 843300,China;Key Laboratory of Livestock and Forage Resources Utilization Around Tarim,Ministry of Agriculture and Rural Affairs,Aral 843300,China;College of Life Science and Technology,Tarim University,Aral 843300,China;Key Laboratory of Biological Resource Conservation and Utilization in Tarim Basin,Xinjiang Production and Construction Corps,Alar 843300,China)

机构地区：[1]塔里木大学动物科学与技术学院,阿拉尔843300 [2]农业农村部环塔里木畜草资源利用重点实验室,阿拉尔843300 [3]塔里木大学生命科学与技术学院,阿拉尔843300 [4]新疆生产建设兵团塔里木盆地生物资源保护与利用重点实验室,阿拉尔843300

出　　处：《中国草食动物科学》2025年第2期17-24,共8页China Herbivore Science

基　　金：国家自然科学基金项目(31860629);塔里木大学研究生科研创新项目(TDGRI202336)。

摘　　要：为探究WNT5B基因在塔里木马鹿茸再生发育中的作用,以塔里木马鹿茸为试验材料,克隆WNT5B基因CDS区序列,利用大肠杆菌重组技术表达塔里木马鹿WNT5B蛋白,并采用生物信息学方法进行序列分析;利用实时荧光定量PCR和免疫荧光检测WNT5B基因在鹿茸不同组织中的表达变化。结果显示,塔里木马鹿WNT5B基因CDS区长1074 bp,编码358个氨基酸;与欧洲马鹿和加拿大马鹿的氨基酸序列相似性达100%,与牛和白尾鹿的相似性为99.4%;塔里木马鹿WNT5B蛋白的二级结构主要以α-螺旋和无规则卷曲为主。在37℃、IPTG浓度1 mM的诱导条件下,3~48 h内成功表达塔里木马鹿茸WNT5B蛋白,分子量约40 kDa。WNT5B基因在塔里木马鹿茸的茸皮、间充质、软骨和骨4个组织中均有表达,其中在骨组织中的表达量最高,极显著高于其他组织(P<0.01);其次在软骨中的表达量极显著高于茸皮和间充质组织(P<0.01)。综上,本研究成功获得了塔里木马鹿茸WNT5B基因CDS区序列,并确定了WNT5B在鹿茸不同组织中的表达特征。In order to investigate the role of WNT5B gene in the regeneration and development of Tarim Red deer antlers,this experiment was conducted using Tarim Red deer antlers as the test material,the sequence of CDS region of WNT5B gene was cloned,and the WNT5B protein of Tarim Red deer was expressed by using the recombinant technology of Escherichia coli and the sequence was analyzed by using bioinformatic methods.The expression changes in different tissues of antlers were detected by real-time fluorescence quantitative PCR and immunofluorescence.The results showed that the CDS region of the WNT5B gene of Tarim Red deer was 1074 bp long,encoding 358 amino acids, with 100% amino acid sequence similarity to Cervus elaphus and Cervus canadensis, and 99.4% similarity tocattle and White-tailed deer. The secondary structure of the WNT5B protein of the Tarim Red deer was mainlybased on the α-helix and irregular coiling. Under the induction temperature of 37°C and IPTG concentration of 1mM, the WNT5B protein of Tarima Red deer antler was successfully expressed within 3-48 hours, with a molecularweight of approximately 40 kDa. The WNT5B gene was expressed in the antler velvet skin, mesenchyme, cartilageand bone of the Tarim Red deer antlers, in which the highest expression was found in the bone tissues, which wassignificantly higher than that in the other tissues (P<0.01), followed by that of cartilage, which was significantlyhigher than that of velvet skin and mesenchyme. In summary, the sequence of the CDS region of WNT5B gene inTarim Red deer was successfully obtained, and the expression characteristics of WNT5B in different tissues of antlerwere determined.

关 键 词：塔里木马鹿 鹿茸 WNT5B基因 分子克隆 免疫荧光 原核表达 

分 类 号：S825.2[农业科学—畜牧学]