title: Construction of a bacterial assay for estrogen detection based on an estrogen-sensitive intein.
abstract: 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 α (hERα). 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 β-galactosidase colorimetric assay. Eight typical chemicals (17-β-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 (EC(50)), 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.affiliation: School of Life Science & Technology, Shanghai Jiao Tong University, 800 Dong-Chuan Road, Shanghai 200240, China.
authors.forename: Rubing
authors.initials: R
authors.lastname: Liang
authors.forename: Jing
authors.initials: J
authors.lastname: Zhou
authors.forename: Jianhua
authors.initials: J
authors.lastname: Liu
mesh_terms: D019677:Artificial Gene Fusion; D001681:Biological Assay / Q000379:methods*; D015374:Biosensing Techniques / Q000379:methods*; D002681:China; D004926:Escherichia coli / Q000378:metabolism*; D047628:Estrogen Receptor alpha / Q000235:genetics / Q000378:metabolism; D004967:Estrogens / Q000032:analysis*; D017930:Genes, Reporter; D006801:Humans; D047668:Inteins; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D014873:Water Pollutants / Q000032:analysis; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
chemical_list: C506487:ESR1 protein, human; D047628:Estrogen Receptor alpha; D004967:Estrogens; D029701:Saccharomyces cerevisiae Proteins; D014873:Water Pollutants; D001616:beta-Galactosidase