BIOLOGY

CONCEPTS & CONNECTIONS

Fourth Edition
Neil A. Campbell Jane B. Reece Lawrence G. Mitchell Martha R. Taylor

CHAPTER 12
DNA Technology and
the Human Genome
Modules 12.1 12.6
From PowerPoint Lectures for Biology: Concepts & Connections
Copyright 2003 Pearson Education, Inc. publishing as Benjamin Cummings

From E.Coli to a Map of Our Genes

Research on E. coli revealed
that these bacteria have a
sexual mechanism that can
bring about the combining of
genes from two different cells
This discovery led to the
development of recombinant DNA technology
asetoftechniquesforcombininggenesfrom
differentsources

Copyright 2003 Pearson Education, Inc. publishing as Benjamin Cummings

 DNA technology has many useful applications

The Human Genome Project
The production of vaccines, cancer drugs, and
pesticides
Engineered
bacteria that
can clean up
toxic wastes

Copyright 2003 Pearson Education, Inc. publishing as Benjamin Cummings

BACTERIA AS TOOLS FOR MANIPULATING

DNA
12.1 In nature, bacteria can transfer DNA in three
ways
Transformation, the taking
up of DNA from the fluid
surrounding the cell

DNA enters
cell

Fragment of
DNA from
another
bacterial cell

Bacterial chromosome
(DNA)
Figure 12.1A
Copyright 2003 Pearson Education, Inc. publishing as Benjamin Cummings

 Transduction, the
transfer of bacterial
genes by a phage

Conjugation, the union

of cells and the DNA
transfer between them
Mating bridge

Phage

Fragment of
DNA from
another
bacterial cell
(former phage
host)

Sex pili

Donor cell
(male)
Figure 12.1B
Copyright 2003 Pearson Education, Inc. publishing as Benjamin Cummings

Figure 12.1C

Recipient cell
(female)

 The transferred DNA is then integrated into the

recipient cells chromosome
Donated DNA

Degraded DNA
Crossovers

Recipient cells
chromosome
Figure 12.1D
Copyright 2003 Pearson Education, Inc. publishing as Benjamin Cummings

Recombinant
chromosome

12.2 Bacterial plasmids can serve as carriers for

gene transfer
An F factor is a DNA
segment in bacteria that
enables conjugation
and contains an origin
of replication

F factor (integrated)
Male (donor) cell
Origin of F replication
Bacterial chromosome
F factor starts
replication and
transfer of chromosome
Recipient cell

Only part of the

chromosome transfers

Figure 12.2A
Copyright 2003 Pearson Education, Inc. publishing as Benjamin Cummings

Recombination can occur

F factor (plasmid)
Male (donor)
cell
Bacterial
chromosome

F factor starts
replication and
transfer

An F factor can exist as a

plasmid, a small circular
DNA molecule separate
from the bacterial
chromosome
Plasmids

Plasmid completes
transfer and
circularizes

Cell now male

Figure 12.2B, C
Copyright 2003 Pearson Education, Inc. publishing as Benjamin Cummings

12.3 Plasmids are used to customize bacteria: An

overview
Plasmids are key tools for DNA technology
Researchers use plasmids to insert genes into
bacteria

Copyright 2003 Pearson Education, Inc. publishing as Benjamin Cummings

Bacterium

Plasmid
isolated

2
3 Gene

Bacterial
chromosome

Cell containing gene

of interest

DNA
isolated

inserted
into plasmid

Plasmid

Gene of
interest

Recombinant DNA
(plasmid)

DNA

Plasmid put into

bacterial cell

Recombinant
bacterium

5 Cell multiplies with

gene of interest

Copies of gene

Gene for pest

resistance
inserted into
plants

Copies of protein

Clones of cell

Gene used to alter bacteria

for cleaning up toxic waste

Protein used to
make snow form
at higher
temperature
Protein used to dissolve blood
clots in heart attack therapy
Figure 12.3

Copyright 2003 Pearson Education, Inc. publishing as Benjamin Cummings

12.4 Enzymes are used to cut and paste DNA

Restriction enzymes
cut DNA at specific
points
DNA ligase pastes
the DNA fragments
together

Restriction enzyme
recognition sequence
1

DNA
Restriction enzyme
cuts the DNA into
fragments

Sticky end
Addition of a DNA
fragment from
another source

Two (or more)

fragments stick
together by
base-pairing

The result is
recombinant DNA

DNA ligase
pastes the strand

Figure 12.4
Copyright 2003 Pearson Education, Inc. publishing as Benjamin Cummings

Recombinant DNA molecule

12.5 Genes can be cloned in recombinant plasmids:

A closer look
Bacteria take the recombinant plasmids and
reproduce
This clones the plasmids and the genes they
carry
Products of the gene can then be harvested

Copyright 2003 Pearson Education, Inc. publishing as Benjamin Cummings

E. coli

1 Isolate DNA

from two sources

Human cell

2 Cut both
Plasmid

DNAs with
the same
restriction
enzyme

DNA

Gene V
Sticky ends

3 Mix the DNAs; they join

by base-pairing

4 Add DNA ligase

to bond the DNA covalently

Recombinant DNA
plasmid

Gene V

5 Put plasmid into bacterium

by transformation

6 Clone the bacterium

Bacterial clone carrying many

copies of the human gene
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Figure 12.5

12.6 Cloned genes can be stored in genomic

libraries
Recombinant DNA
technology allows
the construction of
genomic libraries

Genome cut up
with restriction
enzyme
Recombinant
plasmid
OR

Genomic libraries are

sets of DNA fragments
containing all of an
organisms genes

Copies of DNA fragments

can be stored in a cloned
bacterial plasmid or phage
Copyright 2003 Pearson Education, Inc. publishing as Benjamin Cummings

Bacterial
clone

Plasmid
library

Recombinant
phage DNA

Phage
clone

Phage
library
Figure 12.6