FN Archimer Export Format PT J TI Procarcinogens - Determination and Evaluation by Yeast-Based Biosensor Transformed with Plasmids Incorporating RAD54 Reporter Construct and Cytochrome P450 Genes BT AF NGOC BUI, VAN THU HUYEN NGUYEN, THI THANH MAI, CHI BETTAREL, Yvan YEN HOANG, THI THUY LINH TRINH, THI HAI TRUONG, NAM HA CHU, Hoang THANH THANH NGUYEN, VU DUC NGUYEN, HUU WOELFL, Stefan AS 1:1,2;2:3;3:1;4:4;5:1;6:1;7:1;8:1;9:3;10:5;11:2; FF 1:;2:;3:;4:;5:;6:;7:;8:;9:;10:;11:; C1 VAST, Inst Biotechnol IBT, Natl Key Lab Gene Technol, Hanoi, Vietnam. Heidelberg Univ, IPMB, Heidelberg, Germany. Thai Nguyen Univ, Thai Nguyen Univ Sci, Thai Nguyen, Vietnam. IRD, UMR MARBEC, Montpellier, France. Vietnam Natl Univ Agr, Hanoi, Vietnam. C2 VAST, VIETNAM UNIV HEIDELBERG, GERMANY UNIV THAI NGUYEN, VIETNAM IRD, FRANCE UNIV NATL VIETNAM, VIETNAM UM MARBEC IN DOAJ IF 2.806 TC 8 UR https://archimer.ifremer.fr/doc/00625/73740/74114.pdf LA English DT Article AB 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. PY 2016 PD DEC SO Plos One SN 1932-6203 PU Public Library Science VL 11 IS 12 UT 000392843200087 DI 10.1371/journal.pone.0168721 ID 73740 ER EF