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重组CD200基因在COS-7细胞中的瞬时表达
http://www.100md.com 《中华医学研究杂志》 2006年第12期
CD200;COS-7细胞;,电转染;,流式细胞术,,CD200;COS-7细胞;,电转染;,流式细胞术,1材料与方法,2结果,3讨论,【参考文献
     重组CD200基因在COS-7细胞中的瞬时表达 (pdf)

    【摘要】 目的 研究构建于重组真核表达载体pcDNA3-CD200上的CD200基因在COS-7细胞中的瞬时表达。方法 体外扩增并酶切鉴定pcDNA3-CD200质粒;常规方法培养COS-7细胞;用电转染基因法将pcDNA3-CD200质粒转入COS-7细胞中,用流式细胞术(FCM)检测转染细胞CD200表达。结果 转染COS-7细胞后72h,CD200基因表达阳性的细胞计数率为44.5%。结论 成功的利用电转染基因法将重组真核表达载体pcDNA3-CD200转染到COS-7细胞中,并实现了在COS-7细胞中的瞬时表达,为进一步研究CD200的生物学活性以及信号传导奠定了基础。

    【关键词】 CD200;COS-7细胞; 电转染; 流式细胞术

    Transient expression of recombinant CD200 gene in COS-7 cells

    GAO Song,HAO Bing,HOU Zhi-fu,et al. Department of Clinical Laboratory,Second Affiliated Hospital,Zhejiang University College of Medicine,Hangzhou 310009,China

    【Abstract】 Objective The transient expression of CD200 gene which was constructed to the recombinant eukaryotic expression vector pcDNA3-CD200 was tested. Methods pcDNA3-CD200 plasmid was amplified and tested by an enzyme cutting technique. COS-7 cell was cultured in vitro and transferred with pcDNA3-CD200 by electrotransfer media method. The expression of CD200 was detected by flow cytometry. Results After the COS-7 cell was transferred for 72 hours,the expressing rate was recorded for 44.5%. Conclusion The recombinant eukaryotic expression vector pcDNA3-CD200 was transferred into COS-7 cells successfully by electrotransfer media method. After being transferred to COS-7 cells, the CD200 protein was expressed, which lays a foundation for further studying biological activity and signal transduction of CD200. ......

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