| Expression of HNP1cDNA in CHO-dhfr^- cells |
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| 引用本文： | 刘娟,孙永涛,杜德伟,王临旭,翟嵩,王少杨,汪定成. Expression of HNP1cDNA in CHO-dhfr^- cells[J]. 中国人民解放军军医大学学报, 2004, 19(6): 346-349 |
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| 作者姓名： | 刘娟 孙永涛 杜德伟 王临旭 翟嵩 王少杨 汪定成 |
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| 作者单位： | The Center of Diagnosis and Treatment for Infectious Diseases of PLA，Tangdu Hospital，Fourth Military Medical University，Xi'an 710038，China，The Center of Diagnosis and Treatment for Infectious Diseases of PLA，Tangdu Hospital，Fourth Military Medical University，Xi'an 710038，China，The Center of Diagnosis and Treatment for Infectious Diseases of PLA，Tangdu Hospital，Fourth Military Medical University，Xi'an 710038，China，The Center of Diagnosis and Treatment for Infectious Diseases of PLA，Tangdu Hospital，Fourth Military Medical University，Xi'an 710038，China，The Center of Diagnosis and Treatment for Infectious Diseases of PLA，Tangdu Hospital，Fourth Military Medical University，Xi'an 710038，China，The Center of Diagnosis and Treatment for Infectious Diseases of PLA，Tangdu Hospital，Fourth Military Medical University，Xi'an 710038，China，Department of Clinical Laboratory，Tangdu Hospital，Fourth Military Medical University，Xi'an 710038，China |
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| 摘 要： | ![]()
Objective: To prepare secretary recombinant human neutrophil peptidel (HNP1)and test its antimicrobial activity. Methods: The eukaryotic expression vector pcDNA3. 1/V5-His-TOPO-HNP1 was cotransfected with plasmid pDCH1P11 carrying dhfr gene into dhfr- negative CHO (CHO-dhfr- ) cells and recombinant protein was verified by ELISA; G418 selective medium was used to screen the stably expressing cell clones followed by serial passages in 5×10-8 mol/L and 5×10 mol/L methotrexate (MTX) for gene amplification. Finally 4 cell clones with high expression level were obtained and confirmed by ELISA, RT-PCR and IFA. The bacteriastatic activity of concentrated supernatants was tested in vitro as well. Results: The expression level of recombinant HNPl ranged from 18.85 mg/L·48 h to 47.46 mg/L·48 h per 106 cells that was almost 200-fold increase than that in G418 selective medium. 303 bp segments were amplified from 4 stably tranfec tant clones which matched the length of HNPl cDNA by RT-PCR. Strong fluorescence was
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| 关 键 词： | HNP1cDNA CHO-dhfr^-细胞 基因表达 嗜中性粒细胞 MTX |
Expression of HNPlcDNA in CHO-dhfr~- cells |
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| LIU Juan,SUN Yong-tao,DU De-wei,WANG Lin-xu,ZHAI Song WANG Shao-yang,WANG Ding-cheng The Center of Diagnosis and Treatment for Infectious Diseases of PLA,Tangdu Hospital,Fourth Military Medical University,Xi' an ,China. Expression of HNPlcDNA in CHO-dhfr~- cells[J]. Journal of Medical Colleges of PLA(China), 2004, 19(6): 346-349 |
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| Authors: | LIU Juan SUN Yong-tao DU De-wei WANG Lin-xu ZHAI Song WANG Shao-yang WANG Ding-cheng The Center of Diagnosis Treatment for Infectious Diseases of PLA Tangdu Hospital Fourth Military Medical University Xi' an China |
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| Affiliation: | LIU Juan,SUN Yong-tao,DU De-wei,WANG Lin-xu,ZHAI Song WANG Shao-yang,WANG Ding-cheng The Center of Diagnosis and Treatment for Infectious Diseases of PLA,Tangdu Hospital,Fourth Military Medical University,Xi' an 710038,China Department of Clinical Laboratory,Tangdu Hospital,Fourth Military Medical University,Xi' an 710038,China |
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| Abstract: | ![]()
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| Keywords: | human neutrophil peptidel CHO-dhfr- cells MIX |
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