Biotechnology
Reading quiz
Identify the term that best represents each
description
1. When a bacteria is not affected by chemicals that
interfere with its life processes
2. A rod shaped bacterial cell
3. Chemicals that interfere with bacteria’s life
processes
4. Substance discovered by Alexander Fleming that
can be used to treat bacterial diseases
5. Draw a picture of a typical bacterium and
label with cell wall, pili, nucleoid region
(genetic material)
Identification (“DNA fingerprinting”)
Diagnosing and treating genetic
diseases
Genetically modifying organisms to
make medicines or other useful
products
What Are Common Uses of
Biotechnology?
-
Human Growth Hormone
-
Rice with added vitamins and minerals for
use in third
-
world countries
-
Human tissues grown in animal donors
•
Genetically
Modified
Organisms
–
Pesticide
producing
crops
–
Human
organs in
animals for
transplants
Your Health
Making new or cheaper
drugs
Insulin from bacteria
Antibiotics in milk
•
Testing
for and
treating
genetic
diseases
–
Cancer
–
SCIDS
Genetic Engineering
Selective breeding
Advantages?
Disadvantages?
Increasing variation
Induce mutations
This is the Biotechnology Age
You have been born in it and will live during it
You must be informed in order to make
intelligent decisions
And… you deserve to be qualified to work in an
exciting field
-
as scientists, technicians, receptionists,
business execs, and artists
Prokaryotes
are
identified
by
their
shape,
chemical
nature
of
cell
walls,
movement,
and
how
they
obtain
energy
.
SHAPE
:
Rod
-
bacilli
Spiral
-
spirilla
Spherical
-
cocci
CELL WALLS: Scientists use Gram
staining to tell cell wall type apart.
If the Gram testing is
positive, then the bacteria has thick
peptidoglycan walls
If the Gram testing is
negative, the bacteria has thin walls
inside an outer lipid layers.
MOVEMENT: Flagella
.
Metabolic
Diversity
Identifies
prokaryotes
by
how
they
obtain
energy
.
Heterotrophs
Chemoheterotrophs
-
consume
organic
molecules
for
energy
and
a
supply
of
carbon
.
Photoheterotrophs
-
use
photosynthesis
;
need
organic
compound
like
a
carbon
source
.
Autotrophs
Photoautotrophs
-
use
photosynthesis
EX
:
cyanobacteria
-
Chemoautotrophs
-
make
organic
carbon
molecules
from
CO
2
.
They
obtain
energy
directly
from
chemical
reactions
.
Releasing
energy
-
by
fermentation,
cellular
respiration,
or
both
Obligate
aerobes
-
constantly
need
oxygen
Obligate
anaerobes
-
live
without
oxgen
Facultative
anaerobes
-
live
with/without
oxgen
EX
:
E
.
coli
Life Cycle
Bacteria
can
divide
at
high
speeds,
(every
20
min
.
)
Binary
Fission
-
(asexual)
.
Conjugation
-
Bacteria
exchange
genetic
info
by
forming
a
bridge
between
two
of
them
.
Different
genes
are
transferred
to
each
other
to
increase
genetic
diversity
in
bacteria
populations
.
Transformation
-
To
be
discussed
later
Spore
formation
Antibiotics and resistance
Alexander Fleming and Penicillin
Antibiotics
Antibiotic resistance
Bacteria’s impact on life
Pathogenic bacteria
Toxins
Biowarfare
Production of chemicals and foods
Mining, environmental use
Nitrogen fixation
Decomposers
What Are The Basic Tools Of
Biotechnology?
Micropipets
Gel boxes
Micropipets
Used for accurately measuring very
small amounts of fluids
Used in almost all biotechnology to
measure
1
μ
L = .001 mL = .000001 L = 1 x
10^
-
6 L
2 mL =
μ
L = L
Pipet types
P
-
20 (2
-
20
μ
L) P
-
200 (20
-
200
μ
L) P
-
1000 (200
-
1000
μ
L)
Reading quiz
Viruses contain a coat made up of ____
and their genetic material is _____
Lytic and lysogenic are examples of
what?
How is HIV different than many other
viruses?
The problem…
A disease is causing large pale
spots on the leaves of tobacco
plants
1892, Ivanovski identified the
cause of the disease to be in the
juice
1897, Beijerinck suggested that
tiny particles caused the
disease, and named them
viruses (Latin
-
poison)
Viral structure
Life cycles
HIV and AIDS
HIV is the virus that can lead to AIDS
Transmission
Prevention
AVOID any of the forms of transmission!
HIV mutates, evolves very rapidly
Evades defenses of the immune system
Destroys Helper T cells
Is it living?
Contains genetic material? YEP
Evolves? YEP
Interacts with other organisms? YEP
(parasitic)
Metabolic activity? IN A HOST CELL
Reproduces? IN A HOST CELL
What do you think?
Reading quiz (get SG 3,4C)
Write the term that best represents each
description
1. Circular piece of DNA that replicates
independently of the chromosome(s)
2. Protein that can “cut” DNA at precise spots
3. Purposeful manipulation of DNA
4. Overhanging sequence of bases left after DNA is
“cut”
5. Substance that can be used to identify cells with
our gene of interest
1 point each
1. Circular piece of
DNA that replicates
independently of the
chromosome(s)
2. Protein that can “cut”
DNA at precise spots
3. Purposeful
manipulation of DNA
4. Overhanging
sequence of bases left
after DNA is “cut”
5. Substance that can
be used to identify cells
with our gene of
interest
Plasmid (vector okay)
Restriction enzyme
Genetic engineering
Sticky end
Antibiotic (tetracycline
okay)
Genetic engineering defined:
At a minimum from different species
--
from one bacterial species to another...
Or… as dramatic as DNA from different kingdoms!
Recombination of genetic information by
transferring genetic information from one
organism
to another, usually by way of a recombinant
“vector”
Animal
Plant
Bacteria
Uses in basic research
Simple
:
-
to produce multiple copies of DNA
-
to produce multiple copies of a
protein
More complex
:
-
monitor spread of virus
infecting tobacco plants
Transgenic organisms
Organisms contain
genes from other
organisms
Microorganisms
Animals
Plants
Tools to accomplish genetic engineering
Restriction Enzymes
Protein molecules used to cut DNA
Ligase Enzymes
Protein molecules used to put together DNA
Plasmids
Small DNA molecules used to put new
genes into bacteria
Viruses
Used to put new genes into other living
things
How is DNA cut?
DNA is cut using
restriction enzymes.
Examples for different restriction enzymes:
Eco RI, Hind III
Restriction enzymes cut DNA at specific four or six
-
base
-
pair
-
sequences called
restriction sites.
Examples for different restriction sites:
Eco RI at G AATT C
C TTAA G
Hind III at A AGCT T
T TCGA A
How is a gene inserted?
Gene therapy
1. Identify gene
2. Clone gene
3. Replace the
faulty or missing
gene
4. Result? The
correct protein
should eliminate
cause of the disorder
Advantages
Disadvantages
Transformation
Griffith experiment
Outside DNA gets incorporated into
bacteria
What you have: pGLO plasmid (outside
DNA) and E. Coli (bacteria)
Transformation procedure
How do you get
pGLO into E. Coli?
How do you grow
transformed E. Coli?
How do you identify
transformed cells?
Compentency
CaCl2
Heat/ice shock
Spread E. Coli on
petri dish with agar
and incubate
Amp resistance
Arabinose
GFP protein
pGLO
Origin of plasmid
replication
ON/OFF
switch
Green
Fluorescent
Protein
Beta
-
lactamase
to break down
ampicillin
A plasmid used as a
vector must have
a. An origin of replication
b. A gene conferring resistance
to an antibiotic.
In addition, pGLO
contains
c. The Green Fluorescent
Protein
-
the protein of
interest
d. An ON/OFF switch that
controls the synthesis of GFP by
the presence or absence of the
sugar, arabinose.
Tranformation prelab
Read thoroughly through WB 204
-
206
Make notes on any procedures you don’t
understand. Highlight important information
Use the background information to answer the
following
What is the role of the following items in the
transformation lab and identify each as a gene,
protein, or other:
Ampicillin, GFP, Ampicillin
resistant gene, arabinose, pGLO plasmid, E. Coli,
Interpreting results
LB + DNA :
Prediction, why?
LB + DNA + amp :
Prediction, why?
LB + DNA + amp +
arabinose :
Prediction, why?
Satellite colonies
Lab report
Introduction
Purpose
Deduction
Background info
Materials/procedures
Include amounts
SUMMARIZE procedures (don’t copy from lab)
Results
Data table, picture, bar graph?, written paragraph
Analysis
Support/doesn’t support hypothesis? Interpret results from
each petri dish. Difference between your results and class
average? How can you improve the lab? Importance of lab?
Reciprocal teaching: “Are viruses
alive”
As we’ve done before, take turns reading through the
article (rotate after each paragraph). Each person will
have a different role
Summarizer: Summarizes EACH paragraph read
Predictor: Predict what will happen next
Connector: Discusses connections with class
content/experiences/society
Questioner: Asks under and on the surface questions. Asks if
anyone else has questions
Have your assignment sheet out. I will stamp WB 206
(transformation activity sheet) and give you a stamp
for your RT read based on your discussion
Reading quiz (get SG and transformation activity
stamped, and take out 207)
Write the term the best represents each
description
1. A weakened or killed pathogen
2. A disease when the body launches an
immune response against its own cells
3. The first one was produced by Jenner
using cowpox
4. The body system HIV attacks
Antigen vs. antibody
Vaccines
A weakened or killed virus is injected into the
individual
Elicits an immune response (antibodies)
Does not harm the individual
Gel Boxes
Used to separate
chemicals like DNA
based on size,
charge, etc..
Used for
identification/DNA
fingerprinting,
researching genes,
etc..
Lab B analysis
Voltage vs. current
When was electricity
being conducted? Why?
Electricity allows
electrolysis to occur
Role of phenol red?
Role of buffer?
Requirements for running
DNA through a gel:
Current and a stable pH
H
2
O + NaCl + phenol red + TAE buffer
Note bubbles and swirls
of changing pH
Number
Dye
Charge
1
Brilliant green
2
Janus Green B
3
Methylene Violet
4
Methylene Blue
5
Safranin O
6
Methyl Orange
7
Phenol Red
8
Bromophenol Blue
9
Loading Dye
10
Xylene Cyanol
The Principles of Gel
Electrophoresis
Charge of DNA: ____
If put into an
electric field
, it will move from the _____ to the
____ pole.
A
gel
serves as the porous matrix for the movement of the DNA
molecule.
DNA is cut into
fragments
of different size.
DNA GEL
RFLP (DNA FINGERPRINTING)
Protocol
Polymerase Chain Reaction (PCR)
Purpose: Amplify DNA
quickly
Applications: Acquire
enough DNA to run a DNA
fingerprint
Uses taq polymerase
Mammalian cloning
Nucleus of an
organism’s egg is
destroyed (lets say a
sheep)
Nucleus is removed
of a donor organism
of the same species
and put into the egg
That egg then
develops into a clone
of the donor’s DNA
Issues about cloning
Cloning already happens in nature
(identical twins, bacteria, asexual
reproduction)
What are consequences of
mammalian cloning?
How easy is it to do? Cloning the
sheep Dolly took 277 attempts.
What happens to unsuccessful
eggs?
Currently, there is a ban on human
cloning
Enter the password to open this PDF file:
File name:
-
File size:
-
Title:
-
Author:
-
Subject:
-
Keywords:
-
Creation Date:
-
Modification Date:
-
Creator:
-
PDF Producer:
-
PDF Version:
-
Page Count:
-
Preparing document for printing…
0%
Σχόλια 0
Συνδεθείτε για να κοινοποιήσετε σχόλιο