發(fā)布時間：2018-05-21 05:55

  本文選題：塔里木馬鹿茸 + 血管內(nèi)皮生長因子�。� 參考：《中國畜牧獸醫(yī)》2017年05期

【摘要】：試驗旨在從塔里木馬鹿鹿茸頂端組織中克隆血管內(nèi)皮生長因子(vascular endothelial growth factor,VEGF)的編碼序列,分析其分子特性及其在鹿茸組織中的表達情況。本研究采用改良的Trizol法提取鹿茸頂端組織總RNA,以RT-PCR方法獲得VEGF基因,回收純化后連接到pMD18-T載體,轉化大腸桿菌DH5α感受態(tài)細胞并鑒定,利用免疫組化法確定VEGF蛋白在塔里木馬鹿鹿茸頂端組織茸皮層、間充質(zhì)層和軟骨層中的表達水平。結果顯示,VEGF基因開放讀碼框(ORF)全長為648bp,編碼216個氨基酸。通過其序列比對和進化分析發(fā)現(xiàn),塔里木馬鹿茸中VEGF與人、牛、羊、豬的VEGF核苷酸序列同源性分別為98.75%、96.55%、97.58%和97.56%,其中與人VEGF同源性較高。免疫組化試驗表明,塔里木馬鹿VEGF蛋白在間充質(zhì)、茸皮層和軟骨中均有表達,但無明顯表達差異。說明塔里木馬鹿鹿茸可能會是人類研究再生醫(yī)學和血管相關疾病的理想模型,同時,本研究為不同發(fā)育期塔里木馬鹿鹿茸再生干細胞比較蛋白質(zhì)組學研究提供了基礎試驗數(shù)據(jù)。
[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 偽 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.
【作者單位】： 新疆生產(chǎn)建設兵團塔里木畜牧科技重點實驗室;塔里木大學動物科學學院;
【基金】：兵團塔里木畜牧科技重點實驗室項目(HS201509)
【分類號】：S825

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