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Detecting 1acZ gene expression in living cells with new lipophilic, fluorogenic β ‐galactosidase substrates
Author(s) -
Zhang YuZhong,
Naleway John J.,
Larison Karen D.,
Huang Zhijian,
Haugland Richard P.
Publication year - 1991
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.5.15.1720751
Subject(s) - lac operon , gene , gene expression , plasmid , yeast , microbiology and biotechnology , biology , beta galactosidase , gene product , reporter gene , saccharomyces cerevisiae , chemistry , biochemistry
Current methods for detecting lacZ expression in transformed cells are limited because they require such harsh conditions that viability of the cells after detection is drastically reduced. To overcome this problem, we developed a series of new substrates for detection of lacZ expression in living cells under standard culture or physiological conditions. After incubation with these fluorogenic substrates, cultured lacZ ‐positive mammalian cells appear morphologically normal, continue to divide, and retain the fluorescent product. Because the product is so well retained, fluorescence intensity can be quantitatively related to the level of gene expression. We have demonstrated this correlation using transformed yeast cells bearing various plasmids, each containing the lacZ gene and a unique promoter sequence with known capabilities for promoting gene expression in yeast.— Zhang, Y.‐z.; Naleway, J. J.; Larison, K. D.; Huang, Z.; Haugland, R. P. Detecting lacZ gene expression in living cells with new lipophilic, fluorogenic β ‐galactosidase substrates. FASEB J. 5: 3108‐3113; 1991.