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《Journal of Clinical Otorhinolaryngology Head and Neck Surgery》 2013-07
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Construction of the superantigen SEA transfected laryngocarcinoma cells

JI Xiaobin1 LV Jingli2 LIU Qicai2 XIE Jinghua1(1Department of Otorhinolaryngology,Guangzhou First People′s Hospital,Guangzhou,510180,China;2Experimental Medical Research Center,Guangzhou Medical College)  
Objective:To construct an eukaryotic expression vectors containing superantigen staphylococcal enterotoxin A(SEA)gene,and to identify its expression in laryngeal squamous carcinoma cells.Method:SEA full-length gene fragment was obtained from ATCC13565 genome of the staphylococcus,referencing standard strains producing SEA.Coding sequence of SEA was artificially synthetized.Than,SEA gene fragments was subcloned into eukaryotic expression vector pIRES-EGFP.The recombinant plasmid pSEA-IRES-EGFP was constructed and was transfected to laryngocarcinoma Hep-2 cells.Resistant clones were screened by G418.The expression of SEA in laryngocarcinoma cells was identified with ELISA and RT-PCR method.Result:The subclone of artificially synthetized SEA gene was subclone to eukaryotic expression vector pires-EGFP.Flanking sequence confirmed that SEA sequence was fully identical to the coding sequence of standard staphylococcus strains ATCC13565 in Genbank.After recombinant plasmid transfected to laryngocarcinoma cells,the resistant clones was obtained after screening for two weeks.The clones were selected.The specific gene fragment was obtained by RT-PCR amplification.ELISA assay confirmed that the content of SEA protein in supernatant fluid of cell culture had reached about Pg level.Conclusion:The recombinant eukaryotic expression vector containing superantigen SEA gene is successfully constructed,and is capable of effective expression and continued secretion of SEA protein in laryngocarcinoma Hep-2 cells after recombinant plasmid transfected to laryngocarcinoma cells.
【Fund】: 广东省科技厅产业技术研究与开发资金计划项目(No:2012B031800340);; 广州市科技局社会发展应用基础研究专项(No:2009JI-C501-2)
【CateGory Index】: R739.65
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【Citations】
Chinese Journal Full-text Database 3 Hits
1 JI Xiao-bin,ZANG Lin-quan,XIE Jing-hua,et al.Department of Otorhinolaryngology,Guangzhou First People's Hospital Guangzhou 510180,China;Inhibition of superantigen-activated lymphocytes on the growth of laryngeal carcinoma Hep-2 cells[J];Guangdong Medical Journal;2007-12
2 YUAN Yang1 WANG Xuefeng2 ZHANG Yang3 WANG Jiayu4 LUAN Ning2(1Department of Otolaryngology,Affiliated Hospital of Weifang Medical University,Weifang,261031,China;2Department of Otolaryngology,the First Affiliated Hospital of Liaoning Medical University;3Department of the General Surgery,Peking University Third Hospital;4Department of Otolaryngology,the First Hospital of Harbin);Research of SOCS1 silent DC vaccine on laryngocarcinoma therapy[J];Journal of Clinical Otorhinolaryngology Head and Neck Surgery;2012-04
3 DENG Xiao-fang,ZENG Bo-hang,HU Wei-min,et al.(Guangzhou Tumor Hospital,Guangzhou 510095,China);Construction and Identification of the Superantigen SEA Transfected Hepatocellular Carcinoma Cells[J];Journal of Modern Clinical Medical Bioengineering;2005-05
【Co-citations】
Chinese Journal Full-text Database 7 Hits
1 SUN Jian-cong1,ZENG Bo-hang1,DENG Xiao-fang2 (1 Department of Oncology,Second Affiliated Hospital,Guangzhou Medical College,Guangzhou 510260;2 Tumor Hospital,Guangzhou Medical College,Guangzhou 510095,China);The Expression of SEA and Antigen Presentation Correlated Molecules in SEA Gene Transfected BEL-7402 Hepatocarcinoma Cells[J];Academic Journal of Guangzhou Medical College;2006-06
2 JI Xiaobin 1,LU Jingli 2,CHEN Jingxian 3,LIU Qicai 2,XIE Jinghua 1(1 Department of Otolaryngology,Guangzhou Municipal First People's Hospital,Guangzhou,Guangdong,510180,China;2 Experimental Medicine Research Center,Guangzhou Medical College,Guangzhou,Guangdong,510182,China;3 Department of Pathology and CellBiology,Columbia University,New York,10032,USA);Construction of eukaryotic coexpression vector of recombinant MAGE-A3originating from laryngocarcinoma and Staphylococcal endotoxin A with stable expression of vector in 293T cells[J];Chinese Archives of Otolaryngology-Head and Neck Surgery;2012-07
3 LI Xiu-jing1,YIN Jian-jian1,XIANG Dong-xi2,CHEN Na-na1,ZHENG Cong-long1(1.Department of Pathogeny Biology,Medical College of Dalian University,Dalian 116622,China; 2.School of Medicine,Deakin University,Australia);Research progress in superantigen and its application[J];Journal of Dalian Medical University;2013-01
4 Zhu Damian1 Chen Quan2 Li Yixuan1 Zhu Daoyin2(1Molecular Medicine and Cancer Research Center,Chongqing Medical University,Chongqing 400016;2Department of Immunology,College of Preclinical Medicine,Chongqing Medical University,Chongqing 400016);Construction of Tumor-specific Eukaryotic Expression Vector of sea Gene and Its Function in Human Lung Cancer A549 Cells[J];Biotechnology Bulletin;2009-05
5 JI Xiao-bin,LV Jing-li,CHEN Jing-xian,LIU Qi-cai,XIE Jing-hua.Department of Otorhinolaryngology,Guangzhou Municipal First People′s Hospital,Guangzhou 510180,China;Construction of eukaryotic coexpression vector of recombinant MAGE-A3 originating from laryngocarcinoma and Staphylococcal endotoxin A with stable expression of vector in 293T cells[J];Chinese Journal of Clinicians(Electronic Edition);2012-18
6 HE Hui-xiang1,2,CHEN Yu-qiang1,2,YAN Jiang-hua3,DING Yuan2,WANG Yong-jun2,WANG Sheng-yu3,YE Yan-hua2,XU Ying-yi2,LI Qiao-zhen2 1.Medical College of Nanchang University,Nanchang 330000,Jiangxi Province,China;2.Department of Oncology,NO.174 Hospital of Chinese People's Liberation Army,Xiamen 361003,Fujian Province,China;3.Cancer Research Center,Medical College of Xiamen University,Xiamen 361005,Fujian Province,China;pJAK2 polypeptide,an antagonist of suppressors of cytokine signaling-1,can enhance the antitumor effect of dendritic cells[J];Tumor;2013-06
7 ;Application of gene modified dendritic cells in antitumor immunnotherapy[J];Chinese Journal of Cancer Biotherapy;2013-05
【Secondary Citations】
Chinese Journal Full-text Database 1 Hits
1 JIANG Min~1* QIN Xia~1* HU Qin-hai~1 LI Hai-yan~1 JIANG Shan~1 QIAN Gao-chao~1 LI Ningli~1 QIN Ying~(1,2) (1.Biotherapy Center,Shanghai Institute of Immunology,School of Medicine,Shanghai Jiaotong University,Shanghai 200025,China;2.Institute for Nutrition Science,Shanghai Institutes for Biological Sciences,Chinese Academy of Science,Shanghai 200025,China);Silencing of suppressor of cytokine signaling-1(SOCS1)enhances the therapeutic effect of dendritic cell vaccine in animal model of lung cancer[J];Current Immunology;2007-04
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