Saccharomyces cerevisiae BLYAS, a new Bioluminescent bioreporter for detection of androgenic compounds
Saccharomyces cerevisiae BLYAS, a new Bioluminescent bioreporter for detection of androgenic compounds
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DOI：
10.1128/aem.00589-07
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发表时间：
2007-10-01
影响因子：
4.4
通讯作者：
Sayler, Gary S.
中科院分区：
文献类型：
--
作者：
Eldridge, Melanie L.;Sanseverino, John;Sayler, Gary S.
一株具有自主生物发光能力的酿酒酵母菌被设计成对雄激素化学物质有反应。菌株S. cerevisiae BLYAS在染色体上含有人类雄激素受体，通过在pUTK401上的不同酵母启动子GPD和ADHI之间插入一系列雄激素应答元件，组成性地表达lux4和luxB，从而构建出pUTK420。该质粒与含有醛合成（luxCDE）和FMN还原（frp）所需基因的第二个质粒（pUTK404）共转化，产生了对雄激素化学物质有反应的生物发光生物报告蛋白。以双氢睾酮（DHT）为标准，反应时间为3 ~ 4 h， 50%有效浓度值为（9.7 +/- 4.6）× 10(-9) M。对DHT的检测下限为2.5 × 10(-9) M，对睾酮的检测下限为2.5 × 10(-10) M。由于检测速度快，灵敏度高，且具有自封性，该菌株适合用于具有远程环境监测系统潜力的化学品的高通量筛选。
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 ADHI on pUTK401 that constitutively expressed lux-4 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.