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Biotechnology PowerPoint PPT Presentation


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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

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Biotechnology

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Biotechnology l.jpg

Biotechnology


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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)


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Identification (“DNA fingerprinting”)

What Are Common Uses of Biotechnology?

  • Diagnosing and treating genetic diseases

  • Genetically modifying organisms to make medicines or other useful products

  • - Human Growth Hormone

  • - Rice with added vitamins and minerals for use in third-world countries

- Human tissues grown in animal donors


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Your Health

  • Genetically Modified Organisms

    • Pesticide producing crops

    • Human organs in animals for transplants

  • Testing for and treating genetic diseases

    • Cancer

    • SCIDS

  • Making new or cheaper drugs

    • Insulin from bacteria

    • Antibiotics in milk


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Genetic Engineering

  • Selective breeding

    • Advantages?

    • Disadvantages?

  • Increasing variation

  • Induce mutations


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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


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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.


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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 CO2. 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


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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


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Antibiotics and resistance

  • Alexander Fleming and Penicillin

  • Antibiotics

  • Antibiotic resistance


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Bacteria’s impact on life

  • Pathogenic bacteria

  • Toxins

  • Biowarfare

  • Production of chemicals and foods

  • Mining, environmental use

  • Nitrogen fixation

  • Decomposers


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What Are The Basic Tools Of Biotechnology?

Micropipets

Gel boxes


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Used for accurately measuring very small amounts of fluids

Used in almost all biotechnology to measure

1 μL = .001 mL = .000001 L = 1 x 10^-6 L

2 mL = μ L = L

Micropipets

  • Pipet types

    • P-20 (2-20 μ L) P-200 (20-200 μ L) P-1000 (200-1000 μ L)


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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?


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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)


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Viral structure


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Life cycles


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HIV and AIDS

  • HIV is the virus that can lead to AIDS

  • Transmission

  • Prevention

    • AVOID any of the forms of transmission!


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  • HIV mutates, evolves very rapidly

  • Evades defenses of the immune system

  • Destroys Helper T cells


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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?


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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


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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)

1 point each


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Genetic engineering defined:

Recombination of genetic information by

transferring genetic information from one organism

to another, usually by way of a recombinant “vector”

At a minimum from different species --

from one bacterial species to another...

Or… as dramatic as DNA from different kingdoms!

Bacteria

Plant

Animal


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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


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Transgenic organisms

  • Organisms contain genes from other organisms

    • Microorganisms

    • Animals

    • Plants


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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

Tools to accomplish genetic engineering


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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


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How is a gene inserted?


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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


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Transformation

  • Griffith experiment

  • Outside DNA gets incorporated into bacteria

  • What you have: pGLO plasmid (outside DNA) and E. Coli (bacteria)


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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

Transformation procedure


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A plasmid used as a vector must have

Origin of plasmid replication

ON/OFF switch

a. An origin of replication

b. A gene conferring resistance to an antibiotic.

araC

ori

In addition, pGLO contains

pGLO

c. The Green Fluorescent Protein - the protein of interest

GFP

bla

d. An ON/OFF switch that controls the synthesis of GFP by the presence or absence of the sugar, arabinose.

Beta-lactamase to break down ampicillin

Green Fluorescent Protein


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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,


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Interpreting results

  • LB + DNA : Prediction, why?

  • LB + DNA + amp : Prediction, why?

  • LB + DNA + amp + arabinose : Prediction, why?

  • Satellite colonies


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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?


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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


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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


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Antigen vs. antibody


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Vaccines

  • A weakened or killed virus is injected into the individual

  • Elicits an immune response (antibodies)

  • Does not harm the individual


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Used to separate chemicals like DNA based on size, charge, etc..

Used for identification/DNA fingerprinting, researching genes, etc..

Gel Boxes


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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


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Note bubbles and swirls of changing pH

H2O + NaCl + phenol red + TAE buffer


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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.


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DNA GEL


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libproto

RFLP (DNA FINGERPRINTING) Protocol


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Polymerase Chain Reaction (PCR)

  • Purpose: Amplify DNA quickly

  • Applications: Acquire enough DNA to run a DNA fingerprint

  • Uses taq polymerase


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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


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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


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