HISTOCHEMICAL DETECTION OF A GENE TRANSFERRED BY RETROVIRUS VECTOR IN CULTURED HUMAN KERATINOCYTES

被引:2
作者
GARLICK, JA
ELIAS, J
TAICHMAN, LB
机构
[1] Department of Oral Biology and Pathology, Health Sciences Center, State University of New York, Stony Brook, NY
[2] (School of Dental Medicine), and Department of Pathologyz, Health Sciences Center, State University of New York, Stony Brook, NY
基金
美国国家卫生研究院;
关键词
BETA-GALACTOSIDASE; FEULGEN REACTION; KERATINOCYTES; RETROVIRUS VECTOR;
D O I
10.1179/his.1992.15.4.289
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
The gene encoding beta galactosidase (B-gal), derived from E. coli was transferred by retroviral-mediated transduction to cultured human keratinocytes, and its expression was histochemically detected. Primary cultured keratinocytes were infected with a retrovirus vector carrying the genes for neomycin resistance (neo(r)) and B-gal. Confluent cultures were detached from their dishes, fixed in glutaraldehyde, and stained for expression of B-gal, whose presence was detected by a cytoplasmic blue color. We have modified histochemical staining procedures for B-gal detection to localize this product in cultured epithelial cells. Nuclei were counterstained by the Feulgen reaction, providing visualization of individually stained cells in a multilayered tissue. This technique allows histochemical detection of a gene transferred to cultured keratinocytes and facilitates the study of gene expression and cell fate in these cells.
引用
收藏
页码:289 / 292
页数:4
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