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高效表达Smad2蛋白的A549细胞模型的建立 被引量：4

Establishment of A549 Cell Model Expressing Smad2 Protein Efficiently
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摘要 将携带Smad2-Flag基因的慢病毒包装系列质粒转染293T细胞,进行慢病毒包装。用收集并纯化好的慢病毒感染A549细胞,流式分选筛选出GFP阳性的细胞,用Flag抗体和Smad2抗体进行Western blot鉴定。流式分选得到的GFP阳性的A549细胞经Western blot鉴定能高效表达外源Smad2-Flag融合蛋白,同时空载慢病毒处理的GFP阳性细胞并不表达Smad2-Flag蛋白。结果表明成功构建了高效表达Smad2蛋白的A549细胞系,为进一步研究超级干扰素抗肺癌的分子机制奠定了基础。 This paper conducts lentivirus packaging for lentivirus packaging series plasmid transfection 293T cell carrying Smad2-Flag gene; infects A549 cell with lentivirus collected and purified; screens out GFP positive cells with flow sorting and conducts Western blot evaluation with Flag antibody and Smad2 antibody. Through Western blot evaluation, GFP positive A549 cell obtained by flow sorting can efficiently express exogenous Smad2-Flag fusion protein; meanwhile, GFP positive cell of no-load lentivirus pro- cessing does not express Smad2-Flag protein. The result shows that A549 cell line expressing Smad2 protein efficiently is established successfully, which lays a foundation for further research on anti-lung cancer molecular mechanism of super interferon.
作者 袁素敬 梁余培 陈勇毅 章康健
机构地区 浙江理工大学生命科学学院新元医学与生物技术研究所 中国科学院上海生命科学研究院生物化学与细胞生物学研究所
出处 《浙江理工大学学报（自然科学版）》 2013年第3期372-376,共5页 Journal of Zhejiang Sci-Tech University(Natural Sciences)
基金 浙江理工大学科研启动基金(1116818-Y)
关键词 A549细胞株 慢病毒 GFP SMAD2 A549 cell line lentivirus GFP Smad2
分类号 Q507 [生物学—生物化学]
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参考文献22

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同被引文献22

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引证文献4

二级引证文献9

浙江理工大学学报（自然科学版）
2013年 第3期

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