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08 lac operon
1.
THE lac OPERON ©
2007 Paul Billiet ODWS
2.
The control of
gene expression Each cell in the human contains all the genetic material for the growth and development of a human Some of these genes will be need to be expressed all the time These are the genes that are involved in of vital biochemical processes such as respiration Other genes are not expressed all the time They are switched on an off at need © 2007 Paul Billiet ODWS
3.
Operons An operon is
a group of genes that are transcribed at the same time. They usually control an important biochemical process. They are only found in prokaryotes. © 2007 Paul Billiet ODWS Jacob, Monod & Lwoff © NobelPrize.org
4.
The lac Operon • • • The
lac operon consists of three genes each involved in processing the sugar lactose One of them is the gene for the enzyme βgalactosidase This enzyme hydrolyses lactose into glucose and galactose © 2007 Paul Billiet ODWS
5.
Adapting to the
environment E. coli can use either glucose, which is a monosaccharide, or lactose, which is a disaccharide However, lactose needs to be hydrolysed (digested) first So the bacterium prefers to use glucose when it can © 2007 Paul Billiet ODWS
6.
Four situations are
possible 1. When glucose is present and lactose is absent the E. coli does not produce β-galactosidase. 2. When glucose is present and lactose is present the E. coli does not produce β-galactosidase. 3. When glucose is absent and lactose is absent the E. coli does not produce β-galactosidase. 4. When glucose is absent and lactose is present the E. coli does produce β-galactosidase © 2007 Paul Billiet ODWS
7.
The control of
the lac operon © 2007 Paul Billiet ODWS
8.
1. When lactose
is absent A repressor protein is continuously synthesised. It sits on a sequence of DNA just in front of the lac operon, the Operator site The repressor protein blocks the Promoter site where the RNA polymerase settles before it starts transcribing Repressor protein DNA I O Regulator gene Operator site © 2007 Paul Billiet ODWS RNA polymerase Blocked z y lac operon a
9.
2. When lactose
is present A small amount of a sugar allolactose is formed within the bacterial cell. This fits onto the repressor protein at another active site (allosteric site) This causes the repressor protein to change its shape (a conformational change). It can no longer sit on the operator site. RNA polymerase can now reach its promoter site DNA I © 2007 Paul Billiet ODWS O z y a
10.
2. When lactose
is present A small amount of a sugar allolactose is formed within the bacterial cell. This fits onto the repressor protein at another active site (allosteric site) This causes the repressor protein to change its shape (a conformational change). It can no longer sit on the operator site. RNA polymerase can now reach its promoter site DNA I © 2007 Paul Billiet ODWS O z y Promotor site a
11.
3. When both
glucose and lactose are present This explains how the lac operon is transcribed only when lactose is present. BUT….. this does not explain why the operon is not transcribed when both glucose and lactose are present. © 2007 Paul Billiet ODWS
12.
When glucose and
lactose are present RNA polymerase can sit on the promoter site but it is unstable and it keeps falling off Repressor protein removed RNA polymerase DNA I O z y Promotor site a
13.
4. When glucose
is absent and lactose is present Another protein is needed, an activator protein. This stabilises RNA polymerase. The activator protein only works when glucose is absent In this way E. coli only makes enzymes to metabolise other sugars in the absence of glucose Activator protein steadies the RNA polymerase Transcription DNA I O z y Promotor site © 2007 Paul Billiet ODWS a
14.
Summary Carbohydrates Activator protein Repressor protein RNA polymerase lac Operon +
GLUCOSE + LACTOSE Not bound to DNA Lifted off operator site Keeps falling off promoter site No transcription + GLUCOSE - LACTOSE Not bound to DNA Bound to operator site Blocked by the repressor No transcription - GLUCOSE - LACTOSE Bound to DNA Bound to operator site Blocked by the repressor No transcription - GLUCOSE + LACTOSE Bound to DNA Lifted off Sits on the operator site promoter site © 2007 Paul Billiet ODWS Transcription
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