Journal
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
Volume 77, Issue 7, Pages 2488-2495Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/AEM.02336-10
Keywords
-
Categories
Funding
- National Natural Science Foundation of China [30900010, 30870512]
- Foundation of the Ministry of Education of China [20090073120066]
- Major State Basic Research Development Program of China (973 Program) [2009CB118906]
Ask authors/readers for more resources
Escherichia coli strain DIER was constructed for estrogen detection by inserting an estrogen-sensitive intein (VMA(ER) intein) into the specific site of the constitutively expressed chromosomal lacZ gene. This VMA(ER) intein was generated by replacing the endonuclease region of the Saccharomyces cerevisiae VMA intein with the estrogen binding region of the human estrogen receptor alpha (hER alpha). When there were estrogens or analogs, the splicing of the VMA(ER) intein was induced to produce the mature LacZ protein, which was detected through a beta-galactosidase colorimetric assay. Eight typical chemicals (17-beta-estradiol, bisphenol A, chrysene, 6-OH-chrysene, benz[a] anthracene, pyrene, progesterone, and testosterone) were detected using this DIER strain, and the whole detection procedure was accomplished in 2 h. Their 50% effective concentrations (EC50), relative estrogenic activities, and estradiol equivalency factors were calculated and were quite consistent with those detected with the yeast estrogen screening (YES) system. Furthermore, the estrogenic activities of the synthetic musk samples extracted from the wastewater and waste sludge of a sewage treatment plant of Shanghai (China) were detected, and their results were comparable to those obtained from the YES system and gas chromatography-mass spectrometry (GC-MS). In conclusion, the DIER bioassay could fill a niche for the efficient, rapid, high-throughput screening of estrogenic compounds and has potential for the remote, near-real-time monitoring of environmental estrogens.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available