| 摘要: |
| [目的]检测鹿茸顶端组织GHR基因的表达,为鹿茸再生的分子机理研究提供理论依据.[方法]根据GenBank已发表的牛、山羊和绵羊生长激素受体(GHR)基因序列,用CLUSTAL W软件进行同源性分析,在保守序列设计克隆鹿GHR基因编码序列的引物.以3～4岁健康鹿生长30 d的鹿茸顶端组织提取的总RNA为模板,利用RT-PCR技术克隆GHR基因的cDNA序列;并用原位杂交技术对GHR基因在鹿茸顶端进行组织定位.[结果]成功克隆到了1 967 bp的序列(GenBank登陆号为EU381142),该序列包含GHR基因的全部编码序列.鹿GHR基因与牛、羊、人GHR核苷酸同源性分别为97%,97%和80%,GHR蛋白氨基酸序列同源性分别为97.9%,97.6%和77.3%.杂交结果显示,在皮肤层、间叶细胞层、软骨膜层和软骨层的阳性组均有蓝色杂交信号.[结论]利用RT-PCR技术和原位杂交技术在鹿茸顶端皮肤层、间叶细胞层、软骨膜层和软骨层均检测到GHR的表达. |
| 关键词: 鹿茸 GHR 基因克隆 原位杂交 |
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| 基金项目:西北农林科技大学博士科研启动费（01140501） |
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| cDNA cloning and tissue localization of GHR gene in antler tip |
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| Abstract: |
| 【Objective】 The study was to detect expression of GHR in antler tip to prouide theoretical basis for regeneration of anter molecular machanism.【Method】 A pair of primers were designed according to the homology region of GHR among other species.Total RNA was isolated from antler tip,the growing antlers from 3-4 years spotted deer were removed on the 30th day after the mineralized ones shed,and the cDNA sequence of GHR gene was cloned by RT－PCR technology.Then,in situ hybridization was performed to detect expression of GHR in the antler tip.【Result】 Complete coding sequence of GHR gene was cloned (GenBank accepted number is EU381142),including 1 967 bp.Compared with others,the homology of GHR to bovine,ovine and human was 97%,97% and 80% respectively,but the homology of protein was 97.9%，97.6% and 77.3% respectively.Hybridization signals were obtained using in situ hybridization.【Conclusion】 The results of RT－PCR and in situ hybridization showed that four regions had expression of GHR in deer antler tip. |
| Key words: deer antler GHR gene cloning in situ hybridization |
