Lecture 15

I. Bacteria can undergo change to their DNA in two ways

A. MUTATION
1. Remember the definition from Lecture 8: A
permanent, heritable change in the genetic material.
There is a change in the chemistry of a gene. This
change is perpetuatedIN THE SAME WAY AS
_______________ UNFIXED REPLICATION ERRORS
B. BacterialRECOMBINATION
\\\
1. The process is which a new recombinant
chromosome is formed byONE HELIX
from two organisms. The
genotype of this new chromosome is different
than that of either of the parents. A change in
usually accompanies this
change in genotype.
2. There are three genetic exchange mechanisms
that bacteria use.
i. TRANSFORMATION : A mechanism
where “naked” DNA is taken up by a bacterium.
ii. TRANSDUCTION : A mechanism in
which a bacteriophage carries DNA from a donor
bacterium ot a recipient bacterium.
iii. Conjugation: A mechanism in which DNA is
passed viaTUBE BETWEEN CELLS .
II. Transformation allows bacteria to make a
protein, or proteins, that give them
STRUCTURE that may be beneficial for
their survival. This also allows scientists the
ability to CULTURE a bacterium to perform
certain tasks.
A. In ENVIRONMENT, bacteria can be
transformed with genes that enable them to
DIGEST OIL SPILLS
B. In medicine, bacteria can be
transformed with a gene that enables them to
make INSULIN

III. DNA that bacteria may uptake can be

linear or circular (See How Does It Work?,
right)
A. Linear DNA can TWIST AROUND
ITSELF into the bacterium’s double-
stranded DNA.
B. Circular DNA (plasmids) will not
integrate into host DNA, but instead
RESIDES, where they will be
transcribed and translated to form protein
products. Circular DNA
IV. In today’s experiment, we will transform
bacteria with a constructed plasmid called
pGLO_. The pGLO plasmid has three
important genes:
A. The gene encoding for gfp
1. gfp is the GENE.
2. This gene originates from the
BIOLUMINESCENT JELLYFISH,
Aequorea Victoria.

B. The bla gene

1. Encodes for beta-lactamase,
an ENZYME that will allow
transformed bacteria to be
RESISTANT to antibiotics that have
a beta-lactam ring (e.g. ampicillin).
2. Because of this new
antibioticresistance gene in the
transformed bacteria, we can
SELECT for their growth using a
media that contains ARABINOSE.
 ARAC
ORI

GFP
BLA
 The important genes of a pGLO plasmid:

C. The araC gene

The araC gene 1. Encodes for a ARAC GENE
called regulatory C protein.
In nature, the Regulatory C protein
stops the production of the enzymes
1. The Regulatory C needed to digest the simple sugar
protein (labeled ARABINOSE , if this sugar is
“C”) binds upstream PRESENT in the environment.
of the gene 2. Regulatory C protein binds
encoding for gf . upstream of the arabino se digestive
This blocks RNA
p genes and_____________________
polymerase and ______________. When arabinose is
gfp is not made. present, the Regulatory C protein
2. Arabinose bind to changes conformations and
the Regulatory C TAKES UP the needed genes.
protein.
Conformational 3. In pGLO, the arabinose digestive
changes allow RNA genes_USED_ by
polymerase tobind the gene
___encoding for the green
and transcribe the fluorescent protein. Thus_________
gene encoding for _____________________________
gfp . _____________________________.
V. Methods
A. CaCl 2
1. Often used to render bacterial
membranes moreGENETICALLY
TRANSFORMED.
B. Heat shock
1. CARRY OUT DNA uptake
C. Glucose
1. Allows for quick cell recovery.