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 expression vector of Laryngeal cancer related gene protein

作　　者：杨淑梅 谢海龙[1] 李晓杰[2] YANG Shumei;XIE Hailong;LI Xiaojie(Key Laboratory of Cancer Cellular and Molecular Pathology in Hunan Province,Hengyang 421001,Hunan,China;Cancer Research Institute of Medical College,University of South China)

机构地区：[1]南华大学医学院肿瘤研究所,肿瘤细胞与分子病理学湖南省重点实验室,湖南衡阳421001 [2]郴州市第一人民医院病理科

出　　处：《中南医学科学杂志》2018年第1期32-36,共5页Medical Science Journal of Central South China

基　　金：郴州市科技局资助项目(CZ2013082)

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V152HLCRG1。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。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 9-22,41-52,91-100,103-110,125-134,145-148,151-163,166-177,187-192,209-225,235-242,251-259,273-282 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.

关 键 词：喉癌相关基因 喉癌 抗原表位 重组蛋白 

分 类 号：R3[医药卫生—基础医学]