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

来源：

JOURNAL OF HISTOTECHNOLOGY | 1992年 / 15卷 / 04期

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

页码：289 / 292

页数：4

共 19 条

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