| 冷冻电子显微镜观察人釉原蛋白的自组装 |
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| 引用本文： | 赖婷婷,陈亮,张川,田鲲. 冷冻电子显微镜观察人釉原蛋白的自组装[J]. 实用口腔医学杂志, 2021, 0(1): 114-118 |
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| 作者姓名： | 赖婷婷 陈亮 张川 田鲲 |
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| 作者单位： | 西南医科大学口腔医学院;遵义医科大学口腔医学院;四川省人民医院口腔修复科 |
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| 基金项目： | 四川省科技厅应用基础项目(编号:2018JY0057)。 |
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| 摘 要： | 目的:采用冷冻电子显微镜(cryo TEM)体外观察人釉原蛋白的自组装过程,分析自组装各阶段蛋白的聚集状态及细微结构.方法:提取青少年下颌第三磨牙牙胚细胞总RNA,用RT-PCR获得人釉原蛋白基因全长,再与pMD19-T载体构建重组质粒,转入宿主菌E.coli Top10中诱导表达并纯化.冷冻电子显微镜观察pH值从3....
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| 关 键 词： | 冷冻电子显微镜 釉原蛋白 自组装 |
Observation of human amelogenin self-assembly by cryogenic transmission electron microscopy |
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| LAI Tingting,CHEN Liang,ZHANG Chuan,TIAN Kun. Observation of human amelogenin self-assembly by cryogenic transmission electron microscopy[J]. Journal of Practical Stomatology, 2021, 0(1): 114-118 |
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| Authors: | LAI Tingting CHEN Liang ZHANG Chuan TIAN Kun |
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| Affiliation: | (School of Stomatology,Southwest Medical University,Luzhou 646000,China;School of Stomatology,Zunyi Medical University;Sichuan Provincial People's Hospital,Department of Prosthodontics) |
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| Abstract: | Objective:To observe the self-assembly process of human amelogenin by cryogenic transmission electron microscopy(cryo TEM)in vitro.Methods:Total RNA was extracted from the adolescent's tooth germ of mandibular third molar,and the amelogenin cDNA was obtained from the total RNA through full-length gene of amelogenin by RT-PCR.A plasmid was constructed and inserted into pMD19-T vector,then it transformed into E.coli Top10 to acquire purified proteins.cryo TEM was used to observe and record the self-assembling of amelogenin at pH 3.5 to pH 8.0.Results:When pH=8.0,amelogenin gradually assembled from oligomers into multimers,nanospheres and nanochains within 1-20 min.Conclusion:Under suitable conditions,human amelogenin may self-assemble into nanospheres in stages. |
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| Keywords: | Cryogenic transmission electron microscopy Amelogenin Self-assembly |
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