A flow cytometric method for the rapid detection of beta-galactosidase transfected cells: An in vitro and in vivo study

被引:2
作者
Mouawad, R
Khayat, D
Zerrouqi, A
Ghoumari, AM
Soubrane, C
机构
[1] Lab. of the Med. Oncology Department, Salpetrière Hospital, 75013 Paris
关键词
flow cytometry; beta-galactosidase; transduction;
D O I
10.1016/S0022-1759(97)00032-X
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A flow cytometric method has been developed for the rapid analysis of lacZ transduced cells. The method described is based on an indirect immunofluorescence staining procedure using a monoclonal antibody which binds specifically to P-galactosidase from E. coli and to beta-galactosidase fusion proteins. This technique was used for the quantification in vitro as well as in vivo of beta-galactosidase expression in B16 melanoma cells. The described method is appropriate for a variety of cell types (species, lineage), is simple, quantitative, reliable, rapid and applicable to all constructs containing the lacZ selectable markers. It should prove to be very helpful (1) for the quantification of cells expressing the lacZ reporter gene and (2) for studying gene regulation, including transfection modality, promoter efficacy, enhancer activity, and other regulatory factors.
引用
收藏
页码:51 / 56
页数:6
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