( ficient method for gene transfer. ...)Detection of Reporter Gene Activity in Cell Cultures and Murine Epidermis After Helios Gene Gun-Mediated Particle Bombardment

Fast preparation of many DNA-coated cartridges at one time.

Transformation of many specimens in a short time period.

Here we present data to demonstrate a fast and easy method for transformation of living cells using the Helios gene gun apparatus and the detection of repoter gene activity after particle bombardment under in vitro and in vivo conditions.

Materials and Methods
CELL CULTURE AND ANIMALS
HeLa cells were cultured in minimal essential medium (MEM) supplemented with 10% heat-inactivated fetal calf serum, 104 U of penicillin per ml, and 10 mg of streptomycin per ml on sterile cover slips placed in six-well tissue culture plates until the cells reached approximately 80% confluence.

Inbred male BALB/c mice, 7-9 weeks of age, were used in this study.

PLASMIDS
The plasmid pCMV--gal (Clontech, Palo Alto, CA) was used in experiments demonstrating the expression of -galactosidase gene activity under in vitro and in vivo conditions. The plasmid pCMV without the -galactosidase gene was applied as a negative control. Gold microcarriers, coated with the plasmid pEGFP (Clontech, Palo Alto, CA), were kindly provided by Dr. Rommerskirch, Institute of Biochemistry, Medical Center of the Friedrich Schiller University Jena and were used for the detection of green fluorescent protein (GFP) activity in Helios gene guntransfected HeLa cell cultures.

PREPARATION OF DNA-COATED GOLD MICROCARRIERS
Fifty mg gold microcarriers (diameter: 0.6 or 1.0 m) and 100 l of 0.05 M spermidine were vortexed for 5 sec and sonicated for 10 sec in an ultrasonic bath to break up gold clumps. To obtain a DNA Loading Ratio (DLR) of 2,
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