3.

5 Biotechnology
Essential Idea: Biologists have developed techniques for artificial
manipulation of DNA, cells, and organisms.

Gel Electrophoresis
-Gel electrophoresis is used to separate proteins or fragments of DNA
according to their size
-samples are placed in wells cast in a porous gel
-gel is immersed in a conducting fluid (buffer) and an electric field is
applied
-charged molecules will move through the gel; negative and positive
charges move in opposite directions
-DNA is broken into small fragments; they move faster than the larger ones

Types of GEP:
DNA: southern blot
RNA-northern blot
Protein-western blot

PCR: used to amplify small amounts of DNA

DNA Profiling
-A sample of DNA is obtained, either from a known individual or another
source (eg: crime scene)
-Sequences in the DNA are split into fragments using restriction enzymes
-restriction site: locations in DNA containing short, specific sequences

-restriction enzymes/endonuclease: cut DNA at restriction sites

-Fragments are separated using GEP
-A pattern of bands is produced; this pattern is always the same when taken
from 1 individual
-Individual DNA profiles are compared

Genetic Modification
-Since the genetic code is universal, genes can be transferred from 1 species
to another
-new characteristics can be introduced, such as goats producing milk with
spider silk proteins
-new forms of plants can be produced (eg: production of golden rice)

Techniques to Transfer Genes to Bacteria

Insulin production:
1.
2.
3.
4.
5.

DNA is extracted from human pancreatic cells

Plasmids are isolated from bacteria
Restriction enzymes cut the DNA and plasmids at similar points
Recombinant plasmid is reintroduced to host cell
Bacteria produce insulin which can later be separated and purified

Clones
-Clones are groups of genetically identical organisms derived from a single
original parent cell
-many plant species have natural methods of cloning (eg: a garlic bulb
produces leaves that can produce enough food to grow a group of bulbs
-uncommon, but some animal species also have natural methods of cloning
(eg: hydra clones itself by a process called budding)

Cloning animal embryos:

-In their early stages all cells in an animal are pluripotent
-embryo can divide into 2 or more parts through a process called splitting
or fragmentation

Cloning adult animals using differentiated cells:

-It is relatively easy to clone animal embryos, but at that stage it is
impossible to know whether the embryo will have desirable traits
-it is easy to assess characteristics of adults, but more difficult to clone
them.

-In 2012 biologist John Gurdon won the Nobel Prize for his pioneering
research on cloning
-nuclei from Xenopus tadpoles were implanted into eggs cells without
nuclei, and the eggs developed to form normal tissues of the Xenopus frog

Examples of Artificial Cloning

Rooting of stem cutting:
-Stem cutting are short lengths of stem that are used to clone plants
artificially
-If roots develop from the stem, the cutting can become an independent new
plant
1. Nodes are positions on the stem where leaves are attached. With most
species, the stem is cut below a node
2. Leaves are removed from the lower half of the stem
3. The lowest thirds of cutting is inserted into compost or water, compost
should be sterile and contain plenty of both air and water
4. A clear plastic bag with a few holes prevents excessive water loss from
cuttings inserted into the compost
5. Rooting normally takes a few weeks. Growth of new leaves usually
indicates that the cutting has developed roots

Dolly the Sheep

-The production of Dolly was a pioneering development in animal cloning.
The method that was used is called somatic cell nuclear transfer
1. Adult cells were taken from the udder of an ewe and were grown in a
lab using a medium containing a low concentration of nutrients. This
made genes in the cells inactive so that the pattern of differentiation
was lost.

2. Unfertilized eggs were taken from the ovaries of another ewe. The
nuclei were removed from these cells. Cells from step 1 were fused
with the empty cells in step 2 using a small electric pulse.
3. The embryos were then injected when about 7 days old into the uteri
of other ewes that could act as surrogate mothers. Only 1 of 29
embryos implanted successfully and developed through a normal
gestation