What is an Operon?

Group of genes under control of same promoter Genes have related functions Operons are unique to prokaryotes
Promoter Gene 1 Gene 2 Gene 3

Lac Operon
Why have an operon? operon? How does it work?

Why have an Operon?

Bacteria (Prokaryote)

Ensures coordinated expression of genes Single - On / Off

Promoter

Preferred energy source is glucose If no glucose then must use something else
How is lactose used? Induces synthesis of enzymes that convert lactose to glucose and galactose

Transcription

mRNA
Translation

Lac Operon
Group of genes
Turned on in presence of lactose Encodes proteins that cause breakdown of lactose

What are the functional parts of Lac operon? operon?

Regulatory Gene Promoter Operator B-galactosidase, Permease, Transacetylase

Promoter

Polymerase Transcription Transcription

mRNA
Translation

mRNA
Translation

B-galactosidase

Permease

Transacetylase

B-galactosidase

Permease

Transacetylase

What are the functional parts of an operon? operon?

Regulatory gene Promoter Operator B-galactosidase, Permease, Transacetylase

Lac Operon

Gene transcription control in 2 ways POSITIVE CONTROL = Enhanced gene transcription NEGATIVE CONTROL= CONTROL= Prevents gene transcription

Polymerase

No Gene Transcription

Repressor

Negative Regulation
Lactose Absent
Regulatory Gene

Positive Control
Lactose Present
Regulatory Gene Promoter Operator B-galactosidase, Permease, Transacetylase

Promoter

Operator Weak Transcription

Polymerase Polymerase

No transcription of genes
Repressor protein

Repressor protein

Positive Control
Lactose present
Glucose absent cAMP

What if Lactose and Glucose are both present?

Lactose Present Glucose is Present Regulatory Gene Promoter Operator B-galactosidase, Permease, Transacetylase

Regulatory Gene

Promoter

Operator

B-galactosidase, Permease, Transacetylase

Polymerase

Strong Transcription

Weak Transcription Polymerase

CRP
Repressor protein Repressor protein

What levels are cAMP at? LOW Enhanced Transcription? No.

Review

Your Experiment!
1. 2.

What is the inducer? What does the inducer bind to? Absence of glucose leads to increased levels of what? What does it bind to? How does this lead to enhanced transcription?

Inducer present Enzyme made Enzyme + Substrate = Product Inducer: IPTG ( analog of lactose) Enzyme: B-galactosidase Substrate: BCIG (structural analog of lactose) Product: BLUE

Your Plates B
Substrate Only

Your Plates
Strain A Colour on B Colour on Plate B+I plate Genotype

B+I
Substrate and Inducer

Inducer present Enzyme made Enzyme + Substrate = Product

Streaked 3 different Bacterial strains : Wild type (no mutations) Operator mutant (constitutively on) B-galactosidase mutant (deletion in gene)

B C

Website with animations

This wesite has animations under the Genetics section on the lac operon which will help you if you view these before your lab. http://www.maxanim.com/index.htm http://www.maxanim.com/index.htm