AMES TEST

S E R R A N I L L A , D E N I S A L OU I S E
TATA D , J OH N R OE L
TA N , A N N A PAT R I C I A
V I L L A N U E VA , GA B R I E L L A Y N A
V I VA R , RA P HA E L
Z OL I N A , J A R E N A R I A

INTRODUCTION
The Ames test was developed in the 1970s by
Bruce Ames, Professor of Biochemistry at UC-
Berkeley
A fast and sensitive assay of the ability of a
chemical compound or mixture to induce mutations
in DNA
A positive test indicates that the chemical might act
as a carcinogen
PRINCIPLE
Utilizes specific strains of the bacteria Salmonella
tymphirium to detect mutations
The strains used are known as auxotrophs
Auxotrophs are unable to produce a required
nutrient unless supplied in growth media, in this
case, the amino acid histidine
Ames Test combines a bacterial revertant mutation
assay with a simulation of mammalian metabolism
to produce a highly sensitive test for mutagenic
chemicals in the environment.
A rat liver homogenate is prepared to produce a
metabolically active extract.
The extract is combined with a strain of his-
Salmonella bacteria: in the absence of histidine, the
bacteria are unable to grow on minimal medium
(control result).

Salmonella typhimurium
TA1535 TA1538
They have a mutation in the gene encoding an enzyme
required for histidine biosynthesis
(Histidine Auxotrophs)
Other characteristics of S. typhimurium strains that
enhance their ability to detect mutations:
A mutation distinct from that present in the histidine
biosynthesis gene makes the cell wall of the bacteria more
permeable to large molecules that normally would not enter
the cells.
They also possess a mutation in a gene responsible for
proper excision and repair of DNA damage, thus increasing
their sensitivity to mutagens.
Mutagenicity is the capacity of a chemical or physical agent to
cause permanent genetic alterations.
Mutagens cause changes to the DNA that can affect the
transcription and replication of the DNA, can lead to cell death
Mutagens produces mutations in the DNA, and deleterious
mutation can result in aberrant, impaired or loss of function for
a particular gene, and accumulation of mutations may lead to
cancer.
Origin of Mutagens:
Physical (Ionizing radiations, Ultraviolet radiations, and
Radioactive decay)
Chemical (DNA reactive chemicals such as alkaloids, bromine,
benzene; base analogs; intercalating agents; and metals such
as arsenic, cadmium, and nickel)
Biological (Transposon, Virus, Bacteria)
homogenate and bacterial strain are combined with a suspected mutagenic
substance (X)
induction of revertant colonies indicates that some his- bacteria have
mutated (reverted) to his+ , and therefore that substance X is a
mutagen.
OBJECTIVES
To detect chemically induced mutagenesis.
To determine the mutagenic potential of new chemicals and
drugs because there is a high predictive value.
To determine the ability of a tested substance to cause a
reversal, also called a back-mutation of these auxotrophs to
the original prototrophic state.