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以荧光素酶为报告基因定量检测原型泡沫病毒的指示细胞系的建立 被引量：2

A Quantitative Assay for Measuring of Prototypic Foamy Virus Using a Luciferase-based Indicator Cell Line
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摘要 原型泡沫病毒(prototypic foamy virus,PFV)属于反转录病毒科泡沫病毒亚科.为了在细胞水平定量检测PFV的感染,建立了以萤火虫荧光素酶为报告基因的指示细胞系.在幼小仓鼠肾细胞BHK-21中转染含有PFV LTR启动子及其控制下的荧光素酶报告基因的质粒,经过G418筛选得到稳定整合的细胞系.当PFV感染指示细胞系时,其反式激活因子Tas激活LTR启动子,诱导下游报告基因的表达,通过检测荧光素酶的活性即可对PFV的感染进行定量检测.结果表明,该细胞系可特异指示PFV的感染,相比于传统观察细胞病变的方法,该指示细胞系更为灵敏,是PFV相关研究的有力工具,具有潜在的应用价值. Prototypic foamy virus（PFV）belongs to the Spumaretrovirinae subfamily of the Retroviridae family.In order to quantitate PFV infection in vitro,a PFV indicator cell line （PFVL）was established.Baby hamster kidney cells were transfected with reporter plasmid which contains the firefly luciferase gene driven by a PFV long terminal repeat promoter.The PFV transactivator Tas could activate promoter activity of LTR to express luciferase after PFV infection.Then the infection could be quantified by detection of luciferase activity.PFV indicator cell line could detect PFV infection specifically.Compared with standard assays used to detect PFV infection,the PFVL-based assay is more sensitive than the CPE-based assay.In brief,the PFV indicator cell line is an easy,robust and quantitive method for monitoring PFV infection.
作者 许丰雯 徐丹 李森 王明晓 乔文涛
机构地区 南开大学分子微生物与技术教育部重点实验室 南开大学药学院
出处 《南开大学学报（自然科学版）》 CAS CSCD 北大核心 2011年第3期80-84,共5页 Acta Scientiarum Naturalium Universitatis Nankaiensis
基金 国家自然科学基金(30900068 81071343)
关键词 原型泡沫病毒PFV 荧光素酶报告基因 指示细胞系 prototypic foamy virus（PFV） luciferase indicator cell line
分类号 Q78 [生物学—分子生物学]
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参考文献15

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  • 301sson 0., Escher A., Sandberg G. et al. Engineering of mo- nomeric bacterial luciferases by fusion of luxA and luxB genes in Vibrio harveyi. Gene, 1989,81:335--347.
  • 4Jiang C., Langridge W. H. R., Szalay A. A.. Isolation of de- velopmentally regulated promoter elements from tobacco by T- DNA insertional activation with promoterless luciferase marker gene. In Biol and Chemil Status Re- port. Proceedings of the Vlhh International Symposium on Bi- and Chemiluminescence 1993,Szalay AA, Kric- ka LJ, tanlev P (eds). Chichester: Wiley. 1994:217--221.
  • 5Ozawa K., Sato K., Oh I. eta/. Cell and gene therapy usi- ng mes2 enchymal stem cells (MSCs). Journal of Auttoimmun- iby,2008,30(3): 121-- 127.
  • 6De biol Weger L A., Dunbar : markers rhizosphere. Applied and 57:3641--3644. P., Mahaffee W. F. et al. Use of to detect Pseudomonas spp. in the Environmental Microbiology, 1991,.
  • 7Saeij J. P., Boyle J. P., Grigg M. E. et al. Bioluminescence imaging of Toxoplasma gondii infection in living mice reveals dramatic differences between strains. Infect Immun, 2005, 73 (2):695--702.
  • 8Eishingdrelo H., Cai J., Weissensee P. et a/. A cell-based protein-protein interaction method using a permuted lucifera- se reporter. Current Chemical Genomics, 2011,5:122--128.
  • 9Ishitani M., Xiong L., Stevenson B. et al. Genetic analysis of osmotic and cold stress signal transduction in Arabidopsis: Int- eractions and convergence of abscisic acid-dependent and abs- cisic acid-independent pathways. Plant Ce11,1997,9:935--1949.
  • 10郭文涛,马云云,刘慧涛,李敏,靳静,马晶,赵国强.靶向HIV-1启动子的药物筛选系统的建立[J].郑州大学学报（医学版）,2011,46(1):94-97. 被引量：1

引证文献2

二级引证文献11

南开大学学报（自然科学版）
2011年 第3期

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