1070 l a c Operon
therefore clones which contain inserts can be readily
identified by screening.
See also: Constitutive Expression; lac Operon;
Lederberg, Joshua; Phenotype
lac Operon
J Parker
Copyright ß 2001 Academic Press
doi: 10.1006/rwgn.2001.0738
The lactose or lac operon of Escherichia coli is a cluster
of three structural genes encoding proteins involved in
lactose metabolism and the sites on the DNA involved
in regulation of the operon. The three genes are: (1)
lacZ, which encodes the enzyme b-galactosidase
(which splits lactose into glucose and galactose); (2)
lacY, which encodes lactose permease; and (3) lacA,
which encodes a lactose transacetylase. Functional
b-galactosidase and lactose permease are required for
the utilization of lactose by this bacterium. These
proteins are present in the cell in very low amounts
when the organism is grown on carbon sources other
than lactose. However, the presence of lactose and
related compounds leads to the induction of the
synthesis of these proteins. Interest in understanding
the induction of b-galactosidase by its inducer, lactose,
led Jacques Monod and his associates to begin study-
ing the regulation of lactose metabolism in the 1940s.
These studies were aided by analogs of lactose that
could also be synthesized. Of equal importance, genetic
systems (conjugation and transduction) for E. coli
were known which enabled genetic analysis of mu-
tants with alterations in lactose metabolism.
Throughout the 1950s, Jacques Monod, FrancËois
Jacob, and their colleagues performed physiological
and genetic experiments on lactose metabolism in
E. coli that led to important breakthroughs in our
understanding of gene expression and regulation. It
was found that some inducers were not substrates
of b-galactosidase and some substrates were not
inducers. Elegant genetic experiments involving lac
mutants led in turn to the discovery of regulatory
genes such as lacI, which encoded the lac repressor.
These and other experiments led to the operon model
of gene expression proposed in 1961. The power of
this model was widely appreciated; Jacob and Monod
won the Nobel Prize in 1966.
The genes in an operon are transcribed into a single,
polycistronic messenger RNA (mRNA), in this case
from the lac promoter lacP. The regulatory sites that
are part of the operon also include the lac operator
lacO. When the lactose repressor binds to lacO, a
region immediately upstream of the structural genes
of the lac operon, it prevents transcription of the
operon. This is an example of negative control. In-
ducers of the operon bind to the repressor and cause
a conformational change that leads to the disassocia-
tion of the repressor from the operator. Transcription
of the operon then begins. (Although the gene encod-
ing the lactose repressor is not part of the lac operon,
it is located next to it on the chromosome.)
Later it was discovered that there is another regu-
latory protein, which participates in positive control
of the lac operon. This is the catabolite activator pro-
tein (CAP; also called the cAMP receptor protein,
CRP), which, when bound to cAMP, itself binds to a
region of the lac operon upstream of the promoter and
allows RNA polymerase binding. The CAP protein is
involved in regulation of many operons as part of a
global control system, catabolite repression, which
allows the efficient integration of the metabolism of
different carbon sources.
The E. coli lac operon is of much more than histor-
ical importance. Not only has it proved extremely
useful as a model for studies of gene regulation, it is
also a powerful tool in genetic analysis. For example,
the ease of assaying b-galactosidase, both in vitro
using colorimetric assays and on plates using chromo-
genic substrates, has made lacZ an ideal reporter gene
in a large variety of experimental situations. In addi-
tion, the regulatory system consisting of the lac
repressor and lac operator is often incorporated into
cloning vectors to provide an easily controlled regu-
latory system for cloned genes.
See also: Catabolite Repression; Cloning Vectors;
Induction of Transcription; Jacob, FrancËois; lac
Mutants; Monod, Jacques; Operators; Operon;
Polycistronic mRNA; Promoters; Regulatory
Genes
Lactose
J H Miller
Copyright ß 2001 Academic Press
doi: 10.1006/rwgn.2001.0741
A disaccharide (two sugars joined by an O-glycosidic
bond) commonly found in milk. Lactose is termed a
b-galactoside because it consists of galactose joined
to glucose via a b (1!4) glycosidic linkage. Lactose
is cleaved by the enzyme b-galactosidase to yield
galactose and glucose. The study of the regulation of
b-galactosidase synthesis in bacteria by Jacques Monod