Van Ngoc Bui, Thi Thu Huyen Nguyen, Chi Thanh Mai, Bettarel Yvan, Thi Yen Hoang, Thi Thuy Linh Trinh, Nam Hai Truong, Hoang Ha Chu, Vu Thanh Thanh Nguyen, Huu Duc Nguyen, Wolfl S. (2016). Procarcinogens - Determination and evaluation by yeast-based biosensor transformed with plasmids incorporating RAD54 reporter construct and cytochrome P450 genes. PLoS One, 11 (12), e0168721 [18 p.]. ISSN 1932-6203.
Titre du document
Procarcinogens - Determination and evaluation by yeast-based biosensor transformed with plasmids incorporating RAD54 reporter construct and cytochrome P450 genes
Année de publication
2016
Auteurs
Van Ngoc Bui, Thi Thu Huyen Nguyen, Chi Thanh Mai, Bettarel Yvan, Thi Yen Hoang, Thi Thuy Linh Trinh, Nam Hai Truong, Hoang Ha Chu, Vu Thanh Thanh Nguyen, Huu Duc Nguyen, Wolfl S.
Source
PLoS One, 2016,
11 (12), e0168721 [18 p.] ISSN 1932-6203
In Vietnam, a great number of toxic substances, including carcinogens and procarcinogens, from industrial and agricultural activities, food production, and healthcare services are daily released into the environment. In the present study, we report the development of novel yeast-based biosensor systems to determine both genotoxic carcinogens and procarcinogens by cotransformation with two plasmids. One plasmid is carrying human CPR and CYP (CYP3A4, CYP2B6, or CYP2D6) genes, while the other contains the RAD54-GFP reporter construct. The three resulting coexpression systems bearing both CPR-CYP and RAD54-GFP expression cassettes were designated as CYP3A4/CYP2B6/CYP2D6 + RAD54 systems, respectively and used to detect and evaluate the genotoxic potential of carcinogens and procarcinogens by selective activation and induction of both CPR-CYP and RAD54-GFP expression cassettes in response to DNA damage. Procarcinogens were shown to be predominantly, moderately or not bioactivated by one of the CYP enzymes and thus selectively detected by the specific coexpression system. Aflatoxin B-1 and benzo(a) pyrene were predominantly detected by the CYP3A4 + RAD54 system, while N-nitrosodimethylamine only moderately activated the CYP2B6 + RAD54 reporter system and none of them was identified by the CYP2D6 + RAD54 system. In contrast, the genotoxic carcinogen, methyl methanesulfonate, was detected by all systems. Our yeast-reporter system can be performed in 384-well microplates to provide efficient genotoxicity testing to identify various carcinogenic compounds and reduce chemical consumption to about 53% as compared with existing 96-well genotoxicity bioassays. In association with a liquid handling robot, this platform enables rapid, cost-effective, and high-throughput screening of numerous analytes in a fully automated and continuous manner without the need for user interaction.
Plan de classement
Sciences fondamentales / Techniques d'analyse et de recherche [020]
;
Biotechnologies [084]
Description Géographique
VIET NAM
Localisation
Fonds IRD [F B010068934]
Identifiant IRD
fdi:010068934
