TY - JOUR
T1 - Saccharomyces cerevisiae BLYAS, a new bioluminescent bioreporter for detection of androgenic compounds
AU - Eldridge, Melanie L.
AU - Sanseverino, John
AU - Layton, Alice C.
AU - Easter, James P.
AU - Schultz, T. Wayne
AU - Sayler, Gary S.
PY - 2007/10
Y1 - 2007/10
N2 - 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) × 10-9 M, respectively. The lower limit of detection in response to DHT was 2.5 × 10-9 M, and in response to testosterone it was 2.5 × 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.
AB - 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) × 10-9 M, respectively. The lower limit of detection in response to DHT was 2.5 × 10-9 M, and in response to testosterone it was 2.5 × 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.
UR - http://www.scopus.com/inward/record.url?scp=35148872619&partnerID=8YFLogxK
U2 - 10.1128/AEM.00589-07
DO - 10.1128/AEM.00589-07
M3 - Article
C2 - 17675419
AN - SCOPUS:35148872619
SN - 0099-2240
VL - 73
SP - 6012
EP - 6018
JO - Applied and Environmental Microbiology
JF - Applied and Environmental Microbiology
IS - 19
ER -