DNA Technologies and

Society
What do these
pictures have in
common?

Video on therapeutic
proteins
Video

Moving your gene of interest into a vector or

carrier

12.2 Enzymes are used to cut and paste

DNA

Animation: Restriction Enzymes

Copyright 2009 Pearson Education, Inc.

DNA restriction endonucleasestools of the trade for molecular

technologies

From cloned gene to new host

and product

E. coli bacterium
Plasmid

Bacterial
chromosome

Cell with DNA

containing gene
of interest

Isolate
plasmid

Isolate
DNA

Cut plasmid
with enzyme

DNA
Gene of interest
4

Cut cells DNA

with same enzyme

Gene
of interest

Recombinant
DNA
plasmid

Combine targeted fragment

and plasmid DNA

Add DNA ligase,

which closes
the circle with
covalent bonds

Put plasmid
into bacterium
by transformation

Recombinant
bacterium
8

Clone
of cells

Genes may be inserted

into other organisms

Gene
of interest
7

Animation: Cloning a Gene

Examples of
gene use

Allow bacterium
to reproduce

Genes or proteins
are isolated from the
cloned bacterium

Harvested
Examples of
proteins
protein use
may be
used directly

Eukaryotic genes in prokaryotes

cause problems

Making cDNA
McGraw-Hill Online Learning Center
Play Animation: cDNA
If you want a protein product and
the gene is cloned in a bacterium
you need to use a cDNA
Or clone your gene into a
eukaryotic cell

CRISPR Technology
Many bacteria contain an interesting DNA-based
immune system, called CRISPR.(clustered,
regularly interspaced short palindromic repeats ).
This system consists of a set of DNA-editing
enzymes and an array of repeated sequences
interspersed with DNA that is derived from
pathogens that the bacterium or its ancestors have
been infected with. When these pathogens show
up in the cell, these sequences are used to target
the pathogenic DNA for destruction.
Scientists are exploiting this system to make very
precise changes in DNA of a variety of organisms.
VIDEO

Typical CRISPR region in bacterial genome.

Repeats are important for the functioning of

the system; the spacers contain DNA
sequences derived from pathogens.
The cas genes are concerned with editing
bacterial DNA and cutting foreign DNA.

Interference or Defense Phase of CRISPR System

Adaptation Phase of CRISP System

Attendance exercise
What would you need to engineer the
genes for human growth hormone
into E. coli? You do not need to know
exact details but explain in a brief
narrative. Post answers to the
bulletin board.

CSI-1010

Discovery video:
DNA Forensics

_video backup link_

DNA Profiling

Need to amplify the sample

DNA
Some samples are very small and the
particular region of interest is a small
portion of the DNA. A single hair has
been sufficient to identify a suspect.
If we copy variable regions of the
genome, we would have more DNA for
analysis
The method used to copy these regions
is the polymerase chain reaction or PCR

A close look at PCR

http://www.dnai.org/b/index.html
Manipulation, Techniques, amplifying, PCR
amplification
Manipulation, Techniques, amplifying,
making many copies, amplification graph

Agarose gel electrophoresis to

separate pieces of amplified DNA
by size

Which regions of the DNA are

variable enough to serve as a
fingerprint?
Not genes but the regions in between
genes where there are repeat sequences.
Short tandemly repeated sequences are
highly useful

Animation of STR analysis

http://www.dnai.or
g/d/index.html
Applications,
Human
Identification,
profiling, Todays
DNA profile

Uses of DNA profiling

Guilt or innocence in a crime
Paternity testing
Proved Bill Clinton had sex with Monica
Lewinsky
Thomas Jefferson or a close relative
fathered a son with Sally Hemings, his
slave
Identify victims of mass causalties, like
9/11