Saccharomyces cerevisiae BLYAS, a new bioluminescent bioreporter for detection of androgenic compounds.
A Saccharomyces cerevisiae strain, capable of autonomous bioluminescence, was engineered to respond to androgenic chemicals. The strain, S. cerevisiae BLYAS, contains the human androgen receptor in the chromosome and was constructed by inserting a series of androgen response elements between divergent yeast promoters GPD and ADH1 on pUTK401 that constitutively expressed luxA and luxB to create pUTK420. Cotransformation of this plasmid with a second plasmid (pUTK404), containing the genes required for aldehyde synthesis (luxCDE) and FMN reduction (frp), yielded a bioluminescent bioreporter responsive to androgenic chemicals. Using dihydrotestosterone (DHT) as a standard, the response time and the 50% effective concentration values were 3 to 4 h and (9.7 +/- 4.6) x 10(-9) M, respectively. The lower limit of detection in response to DHT was 2.5 x 10(-9) M, and in response to testosterone it was 2.5 x 10(-10) M. This strain is suitable for high-throughput screening of chemicals with potential for remote environmental monitoring systems because of the assay speed, sensitivity, and self-containment.
The Center for Environmental Biotechnology, The University of Tennessee, 1416 Circle Dr., 676 Dabney Hall, Knoxville, TN 37996-1605, USA.
Melanie L
ML
Eldridge
John
J
Sanseverino
Alice C
AC
Layton
James P
JP
Easter
T Wayne
TW
Schultz
Gary S
GS
Sayler
D000728:Androgens / Q000032:analysis* / Q000378:metabolism; D001681:Biological Assay; D015374:Biosensing Techniques; D017930:Genes, Reporter; D008164:Luminescent Proteins; D010957:Plasmids / Q000235:genetics; D011944:Receptors, Androgen / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D012680:Sensitivity and Specificity; D013997:Time Factors
D000728:Androgens; D008164:Luminescent Proteins; D011944:Receptors, Androgen