Error-prone mutagenesis detected in mammalian cells by a shuttle vector containing the supF gene of Escherichia coli

1984 ◽  
Vol 4 (10) ◽  
pp. 2227-2230
Author(s):  
S Sarkar ◽  
U B Dasgupta ◽  
W C Summers
Keyword(s):  
Escherichia Coli ◽  
Kidney Cell ◽  
Mammalian Cells ◽  
Mutation Frequency ◽  
Shuttle Vector ◽  
Fold Increase ◽  
Host Cells ◽  
Kidney Cell Line ◽  
Monkey Kidney Cell ◽  
Supf Gene

When a shuttle vector containing a tyrosine suppressor tRNA (supF) gene as a target for mutagenesis replicated in a monkey kidney cell line, the frequency of SupF+ mutations was 2.3 +/- 0.5 x 10(-3). When the host cells were treated with ethyl methanesulfonate 40 h before transfection, a 10-fold increase in SupF+ mutation frequency was observed. These results supported the hypothesis that a damage-inducible mutagenic pathway exists in mammalian cells and also demonstrated the utility of this shuttle vector for the study of mutagenesis in mammalian cells.

scholarly journals Error-prone mutagenesis detected in mammalian cells by a shuttle vector containing the supF gene of Escherichia coli.

1984 ◽  
Vol 4 (10) ◽  
pp. 2227-2230 ◽  
Author(s):  
S Sarkar ◽  
U B Dasgupta ◽  
W C Summers
Keyword(s):  
Escherichia Coli ◽  
Kidney Cell ◽  
Mammalian Cells ◽  
Mutation Frequency ◽  
Shuttle Vector ◽  
Fold Increase ◽  
Host Cells ◽  
Kidney Cell Line ◽  
Monkey Kidney Cell ◽  
Supf Gene

When a shuttle vector containing a tyrosine suppressor tRNA (supF) gene as a target for mutagenesis replicated in a monkey kidney cell line, the frequency of SupF+ mutations was 2.3 +/- 0.5 x 10(-3). When the host cells were treated with ethyl methanesulfonate 40 h before transfection, a 10-fold increase in SupF+ mutation frequency was observed. These results supported the hypothesis that a damage-inducible mutagenic pathway exists in mammalian cells and also demonstrated the utility of this shuttle vector for the study of mutagenesis in mammalian cells.

Glycosphingolipids of a green monkey kidney cell line (GMK AH-1)

1982 ◽  
Vol 711 (3) ◽  
pp. 466-477 ◽  
Author(s):  
Jonas Blomberg ◽  
Michael E. Breimer ◽  
Karl-Anders Karlsson
Keyword(s):  
Cell Line ◽  
Kidney Cell ◽  
Monkey Kidney ◽  
Kidney Cell Line ◽  
Monkey Kidney Cell ◽  
Green Monkey

Determination of the spectrum of mutations induced by defined-wavelength solar UVB (313-nm) radiation in mammalian cells by use of a shuttle vector

1988 ◽  
Vol 8 (12) ◽  
pp. 5425-5431
Author(s):  
S M Keyse ◽  
F Amaudruz ◽  
R M Tyrrell
Keyword(s):  
Hot Spots ◽  
Mammalian Cells ◽  
Important Determinant ◽  
Shuttle Vector ◽  
Common Mechanism ◽  
Site Specificity ◽  
Base Pairs ◽  
Dna Photoproducts ◽  
Supf Gene

Mutations induced by UVB (313-nm) radiation, a wavelength in the region of peak effectiveness for sunlight-induced skin cancer in humans, have been analyzed at the sequence level in simian cells by using a plasmid shuttle vector (pZ189). We find that significant differences exist between the types of mutations induced by this solar wavelength and those induced by nonsolar UVC (254-nm) radiation. Compared with 254-nm radiation, 313-nm radiation induces more deletions and insertions in the region sequenced. In addition, although the types of base substitutions induced by the two wavelengths are broadly similar (in both cases, the majority of changes occur at G-C base pairs and the G-C to A-T transition is predominant), an analysis of the distribution of these base changes within the supF gene following irradiation at 313 nm reveals additional hot spots for mutation not seen after irradiation at 254 nm. These hot spots are shown to arise predominantly at sites of mutations involving multiple base changes, a class of mutations which arises more frequently at the longer solar wavelength. Lastly, we observed that most of the sites at which mutational hot spots arise after both UVC and UVB irradiation of the shuttle vector are also sites at which mutations arise spontaneously. Thus, a common mechanism may be involved in determining the site specificity of mutations, in which the DNA structure may be a more important determinant than the positions of DNA photoproducts.

Mutagenesis of a shuttle vector plasmid in mammalian cells

1984 ◽  
Vol 4 (3) ◽  
pp. 435-441
Author(s):  
A Razzaque ◽  
S Chakrabarti ◽  
S Joffee ◽  
M Seidman
Keyword(s):  
Mammalian Cells ◽  
Mutation Frequency ◽  
Shuttle Vector ◽  
Marker Gene ◽  
Point Mutations ◽  
Deletion Mutagenesis ◽  
Bacterial Marker ◽  
Cellular Markers ◽  
Microbiological Procedures ◽  
Insertion Mutants

Recently we and others have reported a high frequency of mutagenesis of shuttle vector plasmids after passage in mammalian cells (Razzaque et al., Proc. Natl. Acad. Sci. U.S.A. 80:3010-3014, 1983; Calos et al., Proc. Natl. Acad. Sci. U.S.A. 80:3015-3019, 1983). The mutation frequency was determined by monitoring the integrity of a bacterial marker gene on the plasmid by standard microbiological procedures. Mutant plasmids contained deletions, insertions of cell DNA, and point mutations. The observed mutation frequency of 1% is much higher than that of cellular markers and could be due to the induction of a mutagenic environment by infection with a replicating plasmid. Alternatively, the hypermutagenesis may be due to some critical transient or persistent difference between the DNA in the plasmid and the cellular chromosome. We performed a number of experiments designed to distinguish between these alternatives, with particular reference to deletion mutagenesis. We conclude that mutagenesis was specific to the plasmid and propose that the majority of the deletion and insertion mutants were generated very early in the infection, before replication of the vector. However, some deletion mutagenesis also occurred after plasmid replication had begun.

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