塔里木马鹿鹿茸VEGF基因克隆及其在不同鹿茸组织中的表达研究

发布时间：2018-05-21 05:55

本文选题：塔里木马鹿茸 + 血管内皮生长因子　；参考：《中国畜牧兽医》2017年05期

【摘要】：试验旨在从塔里木马鹿鹿茸顶端组织中克隆血管内皮生长因子(vascular endothelial growth factor,VEGF)的编码序列,分析其分子特性及其在鹿茸组织中的表达情况。本研究采用改良的Trizol法提取鹿茸顶端组织总RNA,以RT-PCR方法获得VEGF基因,回收纯化后连接到pMD18-T载体,转化大肠杆菌DH5α感受态细胞并鉴定,利用免疫组化法确定VEGF蛋白在塔里木马鹿鹿茸顶端组织茸皮层、间充质层和软骨层中的表达水平。结果显示,VEGF基因开放读码框(ORF)全长为648bp,编码216个氨基酸。通过其序列比对和进化分析发现,塔里木马鹿茸中VEGF与人、牛、羊、猪的VEGF核苷酸序列同源性分别为98.75%、96.55%、97.58%和97.56%,其中与人VEGF同源性较高。免疫组化试验表明,塔里木马鹿VEGF蛋白在间充质、茸皮层和软骨中均有表达,但无明显表达差异。说明塔里木马鹿鹿茸可能会是人类研究再生医学和血管相关疾病的理想模型,同时,本研究为不同发育期塔里木马鹿鹿茸再生干细胞比较蛋白质组学研究提供了基础试验数据。
[Abstract]:The aim of this study was to clone the encoding sequence of vascular endothelial growth factor (endothelial growth) from the tip tissue of Tarim Wapiti deer, and to analyze its molecular characteristics and its expression in the antler tissue. In this study, the total RNAs of pilose antler apical tissue were extracted by modified Trizol method, and the VEGF gene was obtained by RT-PCR method. The VEGF gene was recovered and purified and ligated to pMD18-T vector, and transformed into Escherichia coli DH5 伪 competent cells and identified. Immunohistochemical method was used to determine the expression of VEGF protein in the cortex mesenchymal layer and chondrocytes of Tarim deer antler apical tissue. The results showed that the length of ORF was 648 BP, encoding 216 amino acids. The sequence alignment and evolution analysis showed that the homology of VEGF nucleotide sequence between VEGF and human, cattle, sheep and pig in Tarim velvet antler was 98.7575, 96.55, 97.58% and 97.56%, respectively, and the homology with human VEGF was higher. Immunohistochemical test showed that VEGF protein was expressed in mesenchymal, antler cortex and cartilage of Tarim Wapiti, but there was no significant difference between them. The results suggest that the antler of Tarim Wapiti may be an ideal model for the study of regenerative medicine and vascular related diseases. At the same time, this study provides basic experimental data for the comparative proteomics study of stem cells in different stages of development.
【作者单位】：新疆生产建设兵团塔里木畜牧科技重点实验室;塔里木大学动物科学学院;
【基金】：兵团塔里木畜牧科技重点实验室项目(HS201509)
【分类号】：S825

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