Induction of Mutation of a Synthetic c-Ha-ras Gene Containing Hypoxanthine1
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چکیده
The second base of codon 61 of a synthetic c-l la-ra.vgene was replaced with a hypoxanthine residue in a site-specific manner. Transfection of this gene into NIH3T3 cells by the calcium phosphate procedure resulted in increased focus formation. Total DNA was extracted from transformed cells, and the sequences of the inserted c-l la-ra.v DNA were analyzed by the polymerase chain reaction-single-strand conformation polymorphism method. Mutations with A (or hypoxanthine) to G transition were de tected exclusively. These results suggest that the synthetic c-Ha-nu gene can be used for investigations of mutageneses caused by DNA lesions.
منابع مشابه
Induction of mutation of a synthetic c-Ha-ras gene containing hypoxanthine.
The second base of codon 61 of a synthetic c-Ha-ras gene was replaced with a hypoxanthine residue in a site-specific manner. Transfection of this gene into NIH3T3 cells by the calcium phosphate procedure resulted in increased focus formation. Total DNA was extracted from transformed cells, and the sequences of the inserted c-Ha-ras DNA were analyzed by the polymerase chain reaction-single-stran...
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Synthetic c-Ha-ras genes with an analogue of an abasic site in the first or the second position of codon 12, or in the second position of codon 61 were constructed and transfected into NIH3T3 cells. The genes with the lesions in codon 12 exhibited more focus formation than a normal c-Ha-ras gene, while the gene with the lesion in codon 61 did not. Transformed cells were isolated from the foci, ...
متن کاملc-Ha-ras Containing 8-Hydroxyguanine at Codon 12 Induces Point Mutations at the Modified and Adjacent Positions1
To determine the type of mutation induced by 8-hydroxyguanine in a mammalian system, we examined the mutations induced by a synthetic i -I la-ruv protooncogene containing 8-hydroxyguanine in the second po sition of codon 12 (GGC) in NIH3T3 cells. Transaction of this gene significantly increased the number of transformed foci. The c-Ha-r<w gene present in these foci was analyzed by the polymeras...
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تاریخ انتشار 2006