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《Chinese Journal of Cellular and Molecular Immunology》 2007-01
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Construction and expression of eukaryotic coexpression plasmid containing human MUC1 gene and GM-CSF gene

YUAN Shi-fang~1, SHI Chang-hong~2, YAN Wei~3, YAO Qing~1, LI Nan-lin~1, WANG Ting~1, WANG Ling~1, ZHANG Ying-qi~4 ~1Department of Vascular and Endocrine Surgery, Xijing Hospital; ~2LaboratoryAnimalCenter,~3DepartmentofPathology,~4Biotechnology Center, Fourth Military Medical University, Xi’an 710032, China  
AIM: To construct an eukaryotic coexpression plasmid containing the coding region of human MUC1 tandem repeats gene and GM-CSF gene and to identify its expression in COS-7 cell. METHODS: MUC1 tandem repeats gene was obtained by synthesizing the segments. After identified by restriction endonuclease digestion analysis and DNA sequencing, MUC1 tandem repeats gene and GM-CSF gene were cloned into eukaryotic expression vector pcDNA3.1(+) to construct recombinant plasmid pcDNA 3.1(+)-MUC1-GM-CSF. Then the recombinant plasmid was transfected into COS-7 cell by electroporation and the expression of target gene was detected by immunoflourescence and ELISA. RESULTS: Restriction analysis and DNA sequencing showed that the recombinant plasmid contained the coding region of human MUC1 tandem repeats gene and GM-CSF gene. The expression of MUC1 and GM-CSF was detected. CONCLUSION: The suuessful construction and expression of recombinant plasmid pcDNA3.1(+)-MUC1-GM-CSF lay a foundation for further development of DNA vaccine against breast cancer.
【Fund】: 国家自然科学基金资助项目(30571802);; 陕西省自然科学基金资助项目(2004C2-36)
【CateGory Index】: Q78
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【Citations】
Chinese Journal Full-text Database 2 Hits
1 YUAN Shi fang 1, LI Kai zong 2, WANG Ling 1, YAN Zhen 3, HAN Wei 3, ZHANG Ying qi 3 1 Department of Vascular and Endocrine Surgery, 2Department of Hepato biliary Surgery, Xijing Hospital, 3Biotechnology Center, Fourth Military Medical University, Xi’an 710032, China;Induction of specific cytotoxic T lymphocytes and humoral immune response by MUC1 DNA vaccine in mice[J];Journal of Cellular and Molecular Immunology;2003-04
2 YUAN Shi-fang~1, LI Kai-zong~2, WANG Ling~1, YAN Zhen~3, HAN Wei~3, ZHANG Ying-qi~3~1Department of Vascular and Endocrine Surgery, ~2Department of Hepato-biliary Surgery, Xijing Hospital, ~3Biotechnology Center, Fourth Military Medical University, Xi’an 710032, China;Enhanced inhibitory effect of MUC1 gene vaccine on breast cancer growth by GM-CSF[J];Journal of Cellular and Molecular Immunology;2004-06
【Co-citations】
Chinese Journal Full-text Database 3 Hits
1 YUAN Shi-fang~1, LI Kai-zong~2, WANG Ling~1, YAN Zhen~3, HAN Wei~3, ZHANG Ying-qi~3~1Department of Vascular and Endocrine Surgery, ~2Department of Hepato-biliary Surgery, Xijing Hospital, ~3Biotechnology Center, Fourth Military Medical University, Xi’an 710032, China;Enhanced inhibitory effect of MUC1 gene vaccine on breast cancer growth by GM-CSF[J];Journal of Cellular and Molecular Immunology;2004-06
2 WANG Xue-ju1, WANG Zhi-bo1△, WEI Hong-fei1, WU Xiu-li1, WANG Li2, YU Yong-li2*, WANG Li-ying1*1Department of Molecular Biology, 2Department of Immunology, College of Basic Medicine, Jilin University, Changchun 130021, China;Enhancement of HSP-MUC1 antitumor activity by type C CpG-ODN BW005[J];Chinese Journal of Cellular and Molecular Immunology;2007-04
3 YU Xiao-hui,WANG Jin-hua,WANG Feng(Department of Laboratory,The Second People′s Hospital of Dalian,Dalian 116011 China);Feasibility of Specific Lymphocyte Transformation Test in Appraisal of Vaccinating Effect[J];Chinese Journal of Misdiagnostics;2006-21
【Co-references】
Chinese Journal Full-text Database 10 Hits
1 HUANG Xue-Yong1,2, DUAN Guang-Cai1,2, FAN Qing-Tang2, XI Yuan-Lin1, HUANG Zhi-Gang2, SONG Chun-Hua11Department of Epidemiology, College of Public Health, Zhengzhou University, 2Henan Key Open Laboratory of Molecular Medicine, Zhengzhou 450052,China;Construction and identification of prokaryotic expression system for omp22-hpaA fusion gene from Helicobacter pylori[J];Journal of the Fourth Military Medical University;2007-03
2 ZHANG Lei1,2,CHEN Jian-ping1,2,ZHANG Li1,2,WANG Tao1,LIU Ming-jie1,TIAN Yu1 Department of Parasiotology1,Key Laboratory of Chronobiology of Ministry of Health2,West China Medical Center,Sichuan University,Chengdu 610041,China;Construction of a prokaryotic expression vector carrying mompS-linker-flaA fusion gene and its expression in E.coli[J];Journal of Southern Medical University;2006-12
3 Zhang Wenmin~(1),Jiang Juhong~(2),Zhang Meng~(2),Wen Jianming~(2)1.Division of Pathology,Fujian Medical University,Fuzhou 350004 China2.Division of Pathology,The First Affiliated Hospital,Sun Yat-sen University,Guangzhou 510080 China;Construction and Prokaryotic Expression of Fusion Gene of NY-ESO-1 and Heat Shock Protein 70[J];Journal of Fujian Medical University;2006-06
4 CAO Shao-Xian1,MAO Da-gan2,SHU deng-qun2,CHENG Bao2,WANG Jin-Rong2,LIU Tie-zheng1,YANG Li-Guo2,3(1.Institute of Animal Science,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,P.R.China;2.College of Animal Science & Technology,Nanjing Agricultural University,Nanjing 210095,P.R.China; 3.College of Animal Science & Technology,Huazhong Agricultural University,Wuhan 430070,P.R.China);Construction and Expression of Inhibin DNA Vaccine pEGISI in 293T Cell[J];Acta Ecologiae Animalis Domastici;2007-02
5 SHEN Guo-shun, JIN Ning-yi, QIN Xiao-guang, YIN Rong-lan, ZHUANG Tian-zhong, MA Ming-xiao, ZHENG Min (Key Laboratory of Genetic Engineering, Academy of Military Medical Sciences, Changchun 130062, China);Construction and immuninty of DNA vaccine expressing GP3/GP5 of PRRSV and IL-18 of swine[J];Immunological Journal;2006-06
6 WU Xin,HUANG Zu hu,CHEN Jun1,WU Xin liu,DONG Qing1,LU Bei chuan(Department of Infectious Diseases,the First Affiliated Hospital of NJMU,Nanjin 210029;1Gene Therapy Research Center,the Institute of Infectious Diseases,the 302 Hospital of PLA,Beijing 100;Effects of the Plasmids Expressing Interleukin 12 and 18 on Humoral Immune Responses in H-2~d Mice Induced by DNA Immunization of HBV Core Gene[J];Acta Academiae Medicinae Nanjing;2002-04
7 CHEN Hong-ying1.2,CUI Bao-an1.2,LI Xin-sheng1,XIA Ping-an1,WANG Xue-bin1,FANG Zhong-yi3 (1.College of Animal Husbandry and Veterinary,Henan Agricultural University, Zhengzhou 450002,China;2.Animal Food Safety Key Laboratory of Henan Province,Zhengzhou 450002,China;3.Henan Institute of Veterinary Drug Control ,Zhengzhou 450002,China);Cloning of Fusion Gene Chicken Interleukin-18 and Chicken Interferon-alpha and Construction of Its Prokaryotic Expression Vector[J];Journal of Agricultural Biotechnology;2007-03
8 LI Nan, SUN Yi-Min and ZHAO Bao-Hua~College of life Science, Hebei Normal University, Shijiazhuang 050016, China;Cloning of F Gene of Newcastle Disease Virus HeB02 Isolate and the Study of Its DNA Vaccine[J];Chinese Journal of Biotechnology;2006-03
9 WANG Li-zhe,LEI Lian-cheng,HAN Wen-yu (College of Animal Science and Veterinary Medicine ,Jilin University,Changchun 130062,China);Construction of Prokaryotic Coexpression Vector for Lysis Gene E and Staphylococcus Nuclease Gene and Preparation of E. coli Bacterial Ghost[J];Chinese Journal of Biologicals;2007-08
10 CHENG Chang~1,CHEN Zhen~ 1, 2 ,ZHU Cheng~ 1* (~1State Key Laboratory of Plant Physiology & Biochemistry, College of Life Sciences, Zhejiang University, Hangzhou 310029, China)(~2College of Life Sciences and Pharmaceutical and Chemical Engineering, Taizhou University,Taizhou 317000, China);Construction of plant expression vectors with fusion gene of Helicobacter pylori cagA, ureB and ctb and its genetic transformation in tobacco[J];Acta Microbiologica Sinica;2007-01
【Secondary Citations】
Chinese Journal Full-text Database 6 Hits
1 YUAN Shi Fang 1, LI Kai Zong 2, HAN Wei 3, YAN Zhen 3, ZHANG Ying Qi 3 1Department of Vascular and Endocrine Surgery, 2Department of Hepatobiliary Surgery, Xijing Hospital, 3Biotechnology Center, Fourth Military Medical Universi;Construction of eukaryotic expression vectors containing coding of human full length MUC-1 gene and its expression in COS-7[J];Journal of The Fourth Military Medical University;2002-13
2 YUAN Shi Fang 1, LI Kai Zong 2, YAN Zhen 3, HAN Wei 3, ZHANG Ying Qi 3 1 Department of Vascular & Endocrine Surgery, 2 Department of Hepatobiliary Surgery,Xijing Hospital, 3 Center of Biotechnology, Research Administration, Fourth Military Medical University, Xian 710033, China;Suppressing effect of MUC1 gene vaccine on EMT6 breast cancer growth[J];Journal of The Fourth Military Medical University;2003-04
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4 YANG Xin-wei, SUI Yan-fang, LI Zeng-shan, QU Ping, YE-Jing, WU-Wen , DONG Hai-long, ZHANG Xiu-min Department of Pathology, Fourth Military Medical University, Xi'an 710032, Shaanxi Provine, China;Construction of recombinant SEA gene eukaryotic expression vectors and its induction of immune response against murine hepatoma[J];Journal of Cellular and Molecular Immunology;2002-05
5 YUAN Shi fang 1, LI Kai zong 2, WANG Ling 1, YAN Zhen 3, HAN Wei 3, ZHANG Ying qi 3 1 Department of Vascular and Endocrine Surgery, 2Department of Hepato biliary Surgery, Xijing Hospital, 3Biotechnology Center, Fourth Military Medical University, Xi’an 710032, China;Induction of specific cytotoxic T lymphocytes and humoral immune response by MUC1 DNA vaccine in mice[J];Journal of Cellular and Molecular Immunology;2003-04
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