The FASEB Journal, Vol 5, 3108-3113, Copyright © 1991 by The Federation of American Societies for Experimental Biology

RESEARCH COMMUNICATIONS

Detecting lacZ gene expression in living cells with new lipophilic, fluorogenic beta-galactosidase substrates

YZ Zhang, JJ Naleway, KD Larison, ZJ Huang and RP Haugland
Molecular Probes, Inc., Eugene, Oregon 97402.

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.

This article has been cited by other articles:

				H. Cen, F. Mao, I. Aronchik, R. J. Fuentes, and G. L. Firestone DEVD-NucView488: a novel class of enzyme substrates for real-time detection of caspase-3 activity in live cells FASEB J, July 1, 2008; 22(7): 2243 - 2252. [Abstract] [Full Text] [PDF]

				S.-K. Han, I. M. Abraham, and A. E. Herbison Effect of GABA on GnRH Neurons Switches from Depolarization to Hyperpolarization at Puberty in the Female Mouse Endocrinology, April 1, 2002; 143(4): 1459 - 1466. [Abstract] [Full Text] [PDF]

				B. Fiedler and G. Scheiner-Bobis Transmembrane Topology of alpha - and beta -Subunits of Na+,K+-ATPase Derived from beta -Galactosidase Fusion Proteins Expressed in Yeast J. Biol. Chem., November 15, 1996; 271(46): 29312 - 29320. [Abstract] [Full Text] [PDF]

				T. A. Kohout, J. J. O'Brian, S. T. Gaa, W. J. Lederer, and T. B. Rogers Novel Adenovirus Component System That Transfects Cultured Cardiac Cells With High Efficiency Circ. Res., June 1, 1996; 78(6): 971 - 977. [Abstract] [Full Text]

				J. M. Metzger, W.-I. Lin, and L. C. Samuelson Vital Staining of Cardiac Myocytes During Embryonic Stem Cell Cardiogenesis In Vitro Circ. Res., April 1, 1996; 78(4): 547 - 552. [Abstract] [Full Text]

				C. Gillis, A. Haegerstrand, B. Ragnarson, and L. Bengtsson Rapid visualization of viable and nonviable endothelium on cardiovascular prosthetic surfaces by means of fluorescent dyes J. Thorac. Cardiovasc. Surg., December 1, 1994; 108(6): 1043 - 1048. [Abstract] [Full Text]

				J. Frangioni, N Moghal, A Stuart-Tilley, B. Neel, and S. Alper The DNA binding domain of retinoic acid receptor beta is required for ligand-dependent suppression of proliferation. Application of general purpose mammalian coexpression vectors J. Cell Sci., January 4, 1994; 107(4): 827 - 838. [Abstract] [PDF]