| Mutagenesis detection experiments are a kind of experiment used for detecting genetoxic substance. At present, the category of experiment detected mutagen are as much as 200, shuttle plasmid/ mammalian cell mutagenesis detection system can reveal the mutation mechanism of higher animal veridically, because mutation occur in the cell of mammalian cell. Moreover, single base replacement at any one location can be detected through the change of phenotypic, sensitivity is high. As the experiment operation of it is simple and the outcome is exact the detection system are applied generally in the screening of mutagen in foods and environment and the mutagenesis research of various kinds of mutagens.We transfection shuttle plasmid pSP189 to Vero cell of mammal, after cultivating for 4872h,reclaiming plasmid, transformating to host bacterium MBM7070,screening muton and measuring spontaneous mutation of it ,the outcome is 2.360×10-4,between 2.5×10-4 and 1.8×10-4 Reported by literature, meeting the request that spontaneous mutation frequency of mutation detection system is low. Establishing successfully the cell mutagenesis detection system of shuttle plasmid pSP189/ mammal Vero cell.For verifying if the system established can be used for the detection of mutagen, we utilize direct positive Mu MNNG and indirect positive Mu B(a)p which have be known to verify the system., the concentration of MNNG to use narcotics to plasmid is 0.32μg/ml,experiment undertakes four parallel, at the same time committing four group DMSO solvent as control group. Result display the mutation frequency of MNNG group is 19.6×10-4 , and control group is 2.1×10-4. the mutation frequency of MNNG group is 9.3 times solvent control groups, mutation detection is obvious, according to this, the system can be used to detect the direct acting mutagens. The concentration of B(a)p to use narcotics to plasmid is 1.1μg/ml, experiment undertakes four parallel, committing four group DMSO solvent as control group meanwhile.
|
PDF Full Text Request