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Visualizing interleukin 2 gene expression at the single cell level.

作者信息

Emilie D, Peuchmaur M, Barad M, Jouin H, Maillot M C, Couez D, Nicolas J F, Malissen B

机构信息

Centre d'Immunologie INSERM-CNRS de Marseille-Luminy, France.

出版信息

Eur J Immunol. 1989 Sep;19(9):1619-24. doi: 10.1002/eji.1830190915.

Abstract

To analyze the expression of the interleukin (IL)2 gene at the single cell level, we have constructed a chimeric gene in which the regulatory sequences from the mouse IL2 gene were fused 5' proximal to the coding region of the Escherichia coli beta-galactosidase (lacZ) gene. Once stably introduced into a T cell hybridoma, the IL 2-lacZ reporter gene was shown to display a pattern of induction similar to the one observed for the resident IL 2 gene. Two points emerged from monitoring IL 2 expression for the resident IL2 gene. Two points emerged from monitoring IL 2 expression at the single cell level. First, upon activation the expression of the IL2-lacZ gene appears asynchronous. Second, at the peak of induction the percentage of beta-galactosidase+ cells never reached 100% and the level of beta-galactosidase reached among positive cells was highly heterogeneous within a cloned population of T cells. In combination with the possibility to sort viable lymphocytes according to lacZ expression, the IL2-lacZ reporter gene described herein should provide a way to isolate somatic cell mutants deficient in signal transduction by the T cell antigen receptor complex.

摘要

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