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We describe a method to identify and enumerate mutants at the nucleotide level in complex cell populations. Several thousand different mutants were induced at the HPRT locus in human lymphoblastoid cultures by either MNNG, an alkylating agent, or by ICR-191, a substituted acridine. HPRT mutants were selected en masse by resistance to 6-thioguanine. The most frequent mutations (hotspots) in HPRT exon 3 were determined by a combination of denaturing gradient gel electrophoresis and polymerase chain reaction. MNNG predominantly produced GC----AT transitions at nucleotides in a GGGGGG sequence, while ICR-191 produced both +1 frameshifts in the same GGGGGG sequence and +1 frameshifts in a CCC sequence.
N F Cariello, and
P Keohavong, and
A G Kat, and
W G Thilly
January 2020,
Methods in molecular biology (Clifton, N.J.),
N F Cariello, and
P Keohavong, and
A G Kat, and
W G Thilly
January 1994,
Environmental and molecular mutagenesis,
N F Cariello, and
P Keohavong, and
A G Kat, and
W G Thilly
March 1978,
Mutation research,
N F Cariello, and
P Keohavong, and
A G Kat, and
W G Thilly
May 2000,
Mutation research,
N F Cariello, and
P Keohavong, and
A G Kat, and
W G Thilly
November 1981,
Journal of molecular biology,
N F Cariello, and
P Keohavong, and
A G Kat, and
W G Thilly
February 1977,
Mutation research,
N F Cariello, and
P Keohavong, and
A G Kat, and
W G Thilly
June 2009,
Human genetics,
N F Cariello, and
P Keohavong, and
A G Kat, and
W G Thilly
April 1986,
Mutation research,
N F Cariello, and
P Keohavong, and
A G Kat, and
W G Thilly
November 2000,
Genetics,
N F Cariello, and
P Keohavong, and
A G Kat, and
W G Thilly
May 1980,
Mutation research,
