Transfection of DNA

into Eukaryotes
Mom, please, may I have a
glowing fish, too?

Although glowing fish are

fun, the procedure used to
make them is very useful to
scientists!

We are going to make

living cells glow

Transfection
Delivery of foreign molecules (DNA)
into eukaryotic cells
Cell begins to read the piece of DNA
and produce the protein encoded on it

How can we get DNA into a

cell?
Inject it
Poke holes in the membrane
without destroying the cells
Create a complex of DNA plus
some substance that will go into
the cell

We will use Cationic

liposomes
It creates an artificial
membrane
Membrane formed
around pieces of DNA
Cell membrane will fuse
with this artificial
membrane and bring the
DNA into the cell
Liposomes are also be useful
for drug and cosmetic delivery

 Liposome-DNA complex taken up by

the cell by fusing with the cell
membrane

 Can change an
entire animal or just
a culture of cells
View with
fluorescent
microscope
Use blue or
ultraviolet light

GFP
Green fluorescent protein
Gives luminescent jelly fish their greenish
glow

 GFP is a marker protein

on a plasmid vector
A plasmid is a piece of DNA
coding for a specific protein
If the cell read the DNA and
produces the protein, it will
also produce GFP.
That particular protein will
glow
If there is enough of that
protein, the entire cell will
glow

Plasmid
Vector

What can we do with

glowing cells?

Determine distribution of protein

Track movement of protein
Track movement of cells
VERY useful for scientists!