| HSV-1gC糖蛋白真核表达载体的构建及表达 |
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| 引用本文： | 党引利,阎岩,张晓晓,李璞媛,于澜,张蕾,张芳琳,徐志凯,吴兴安. HSV-1gC糖蛋白真核表达载体的构建及表达[J]. 细胞与分子免疫学杂志, 2011, 27(5) |
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| 作者姓名： | 党引利 阎岩 张晓晓 李璞媛 于澜 张蕾 张芳琳 徐志凯 吴兴安 |
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| 作者单位： | 第四军医大学基础部微生物学教研室,陕西,西安,710032 |
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| 摘 要： | 目的:构建单纯疱疹病毒1型(HSV-1)囊膜糖蛋白C(gC)哺乳动物表达载体pCI-mCMV-gC-1-IRES-DHFR-L22R,实现其在中国仓鼠卵巢细胞(CHO-K1)中的稳定表达.方法:利用基因重组技术构建同时克隆有gC-1基因和中国仓鼠二氢叶酸还原(DHFR-L22R)基因的真核表达载体pCI-mCMV-gC-1-IRES-DHFR-L22R,按Lipofectamine(TM)2000说明书转染野生型CHO-K1细胞,在G418和氨甲喋呤(MTX)双筛选压力下筛选稳定表达细胞株,Slot blot方法测定重组蛋白的表达,并用His-Ni琼脂糖填料进行目的蛋白纯化,纯化后West-ern blot检测.结果:成功构建了真核表达载体pCI-mCMV-gC-1-IRES-DHFR-L22R.经过两轮筛选获得稳定表达HSV-1型邪基因的细胞系,Western blot成功检测到目的蛋白.结论:HSV-1 gC在CHO-K1细胞中成功表达,并具有良好的特异性结合活性,为进一步的实验研究和临床应用提供了基础.
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| 关 键 词： | 真核表达 |
Construction and expression of the eukaryotic expression vector carrying HSV-1 gC glycoprotein gene |
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| DANG Yin-li,YAN Yan,ZHANG Xiao-xiao,LI Pu-yuan,YU Lan,ZHANG Lei,ZHANG Fang-lin,XU Zhi-kai,WU Xing-an. Construction and expression of the eukaryotic expression vector carrying HSV-1 gC glycoprotein gene[J]. Chinese journal of cellular and molecular immunology, 2011, 27(5) |
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| Authors: | DANG Yin-li YAN Yan ZHANG Xiao-xiao LI Pu-yuan YU Lan ZHANG Lei ZHANG Fang-lin XU Zhi-kai WU Xing-an |
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| Affiliation: | DANG Yin-li,YAN Yan,ZHANG Xiao-xiao,LI Pu-yuan,YU Lan,ZHANG Lei,ZHANG Fang-lin,XU Zhi-kai,WU Xing-an Department of Microbiology,Fourth Military Medical University,Xi'an 710032,China |
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| Abstract: | AIM: To stably express herpes simplex virus type 1(HSV-1) glycoprotein C(gC) in Chinese hamster ovary cells(CHO-K1).METHODS: The eukaryotic expression vector pCI-mCMV-gC-1-IRES-DHFR-L22R was constructed and transfected into CHO-K1 cells by LipofectamineTM 2000.The transfected cells were selected by G418 and methotrexate(MTX).The expression of HSV-1 gC was analyzed by Slot blot.HSV-1 gC proteins were purified with His-Ni Sepharose and then detected by Western blot.RESULTS: The eukaryotic expression vector pC... |
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| Keywords: | HSV gC DHFR CHO |
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