| 杨淑梅,谢海龙,李晓杰.喉癌相关基因的生物信息学分析及真核表达载体的构建.[J].中南医学科学杂志.,2018,(1):32-36. |
| 喉癌相关基因的生物信息学分析及真核表达载体的构建 |
| Bioinformatics analysis and construction of eukaryotic expressionvector of Laryngeal cancer related gene protein |
| 投稿时间：2017-08-03 修订日期：2017-11-29 |
| DOI：10.15972/j.cnki.43-1509/r.2018.01.008 |
| 中文关键词: 喉癌相关基因 喉癌 抗原表位 重组蛋白 |
| 英文关键词:Laryngeal carcinoma related gene Laryngeal carcinoma epitope recombinant protein |
| 基金项目:郴州市科技局资助项目(CZ2013082). |
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| 摘要点击次数: 658 |
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| 中文摘要: |
| 喉癌相关基因(LCRG1)是喉癌候选抑瘤基因,目前能用于临床病理标本免疫组化的单克隆抗体依然甚少。本研究通过生物信息学分析喉癌相关基因蛋白的二级结构及抗原表位,得出最有可能的抗原表位区域为9-22,41-52,91-100,103-110,125-134,145-148,151-163,166-177,187-192,209-225,235-242,251-259,273-282等13个区域。利用PCR扩增喉癌相关基因,获得867 bp,将该片段插入真核载体V152H中,构建真核表达质粒V152H-LCRG1。再将该质粒转染到人胚肾HEK293细胞,经SDS-PAGE证实成功表达后用Ni2＋2NTA柱层析纯化,得到纯度＞95%的蛋白,为筛选单克隆抗体奠定了理论基础。 |
| 英文摘要: |
| Laryngeal cancer related gene (LCRG1) is a candidate tumor suppressor gene of laryngeal carcinoma.At present,there are few monoclonal antibodies that can be used for immunohistochemistry of clinicopathological specimens.This study analyzed the secondary structure and epitope of LCRG1 protein throμgh bioinformatics,and obtained that the most likely epitope region is 9-22,1-52,1-100,3-110,5-134,5-148,1-163,6-177,7-192,9-225,5-242,1-259,3-282 et al.The LCRG1 fragment was amplified by PCR,and 867bp was obtained.The fragment was inserted into eukaryotic vector V152H,to construct eukaryotic expression plasmid V152H-LCRG1.The plasmid was transfected into human embryonic kidney HEK293 cell.After successful expression by SDS-PAGE verified,the plasmid was purified by Ni2+2NTA column chromatography to obtain protein with purity>95%,which laid a theoretical foundation for screening monoclonal antibodies. |
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