BIO 103: Intro to Biotechnology
What % of pregnancies in the US are assisted by biotechnology?
1% of all births in the US are assisted.
What is E. coli?
A bacterium commonly found in the intestines of humans and other animals, some strains of which can cause severe food poisoning.
What is an antibiotic?
A medicine (such as penicillin or its derivatives) that inhibits the growth of or destroys microorganisms.
What is Arabinose?
A monosaccharide that activates the promoter in the Arabinose operon.
What is the CaMV 35S Promoter?
A promoter from the genome of the cauliflower mosaic virus. 35S is a strong promoter which means that it causes a high level of gene expression.
What is Green Fluorescent Protein? (Where do they come from and why are they useful in biology?)
A protein that exhibits green fluorescence when exposed to blue light, specifically that originally isolated from the jellyfish Aequorea victoria, often used as an experimental tracer or marker, especially in molecular biology and genetic engineering
What is Tubulin?
A protein that makes up the microtubules in the cytoskeleton in a cell. A structural protein that is present in all plants.
What is Ampicillin?
A semi-synthetic form of penicillin used to treat infections of the urinary and respiratory tracts.
What is a plasmid?
A small circular form of DNA that is naturally found in some bacteria, but not required for the survival of cells under normal conditions.
What is Polymerase Chain Reaction (PCR)?
A technique used to amplify, or make many copies, of a specific segment of DNA. Uses a machine called a thermal cycler. DNA template, DNA primers, nucleotides, and DNA polymerase are all necessary components of a PCR reaction.
What is a Gene?
A unit of heredity which is transferred from a parent to offspring and is held to determine some characteristic of the offspring.
How are IPS cells made?
Adult cells are collected and grown in culture. Then they are treated with IPS reprogramming factors which causes the adult cells to change into a pluripotent state. These cells can then be treated to cause differentiation into whatever type of body cell is needed.
What is the difference between adult and embryonic stem cells?
Adult stem cells are multipotent. Embryonic stem cells are totipotent and pluripotent.
What are Transgenic animals?
An animal that contains genes from another species. i.e. transgenic mice: can't be affected by polio virus, but when the human cell that serves as a receptor for polio is inserted into the mice's genes, they can be affected by polio.
What is Genetically Modified Organism (GMO)?
Any organisms that has been altered to contain DNA from another organism as part of its genome.
What are stem cells?
Are unspecialized cells that have the potential to give rise to many kids of specialized cells. Used to repair and renew the body. Are considered immortal (can divide indefinitely).
Problems associated with cloning Dolly?
At age 1, DNA was "older" than normal 1 year old sheep. Diagnosed with arthritis at 5 years old. Diagnosed with lung cancer at 7 years old. Died 6 and 1/2 years earlier than a normal sheep.
How does PCR compare/contrast with DNA replication within a cell?
Both use DNA as the template molecule for the reaction. PCR: reaction occurs in vitro; requires high heat to separate DNA strands. DNA Replication: requires sort RNA primers.
Why is the heat shock an important part of the transformation procedure? a) Heat may help denature the bacteria's DNA polymerase. b) Heat may help kill bacteria that don't pick up plasmid. c) Heat may help bacteria keep the plasmid that they have picked up. d) Heat may help unzip the plasmid.
c) Heat may help bacteria keep the plasmid that they have picked up.
In a PCR experiment, the [blank] allows us to amplify only a specific target sequence of DNA because it is complementary to our gene of interest. a) Enzyme. b) PCR bead. c) Primer. d) Instagene.
c) Primer.
Which enzyme is used in Polymerase Chain Reaction to add nucleotides to the new DNA strand? a) Helicase. b) Human DNA Polymerase. c) Taq DNA Polymerase. d) Restriction Enzyme BamH1.
c) Taq DNA Polymerase.
What was the purpose of putting the E. coli without plasmid sample onto an LB + Amp plate? a) To prove that the E. coli - plasmids were resistant to antibiotic. b) To prove that the E. coli + plasmids produced GFP protein. c) To prove that the Ampicillin was effective. d) To prove that the LB contained nutrients.
c) To prove that the Ampicillin was effective.
If you performed the experiment correctly, you would expect: a) colonies of bacteria on the LB + Amp plate for the E. coli - plasmids sample. b) no bacteria on the LB plate for the E. coli - plasmids sample. c) colonies of bacteria on the LB + Amp plate for the E. coli + plasmids sample. d) no bacteria on the LB plate for the E. coli + plasmids sample.
c) colonies of bacteria on the LB + Amp plate for the E. coli + plasmids sample.
The gene for GFP on the plasmid, pGLO, is regulated by: a) the origin of replication. b) the Ampicillin resistance gene. c) the Arabinose promoter. d) the gene for GFP is not regulated.
c) the Arabinose promoter.
A change in pheotype of the E. coli occurs only when: a) the E. coli takes up new plasmid DNA into the cell. b) the newly acquired plasmids are replicated and passed onto offspring. c) the newly acquired plasmid genes are transcribed and translated into protein. d) the transformed E. coli die from the antibiotic.
c) the newly acquired plasmid genes are transcribed and translated into protein.
What are differentiated cells?
Cells that are specialized for some function of the body. Have a limited life span. i.e. skin cells, brain cells, etc.
What are Totipotent Stem Cells?
Cells with the potential to become all types of cells. Capable of diving and developing to form a complete, mature organism.
What are Pluripotent Stem Cells?
Cells with the potential to become almost all types of cells. Capable of developing into many different cell types i.e. muscle cells, nerve cells, bone cells, etc.
What are Multipotent Stem Cells?
Cells with the potential to become more than one type of cell. i.e. blood stem cells can become red blood cells or white blood cells.
What is the difference between changing an organism's genotype (the genetic makeup) and its phenotype?
Changing genotype is changing the genetic makeup (DNA) whereas its phenotype is the physical trait that is expressed.
Reproductive Cloning
Creating new individuals or animals.
Who is Dolly?
Dolly the Sheep. First cloned mammal from adult cells in the 1990s.
What is the controversy with using embryonic stem cells?
Embryo cells come from aborted pregnancies. Playing "God"
Where do scientists obtain (sources) embryonic stem cells vs. adult stem cells?
Embryonic Stem Cells: from the embyro. Adult Stem Cells: bone marrow, spinal cord.
LB + AMP + ARA Plate: Bacteria + plasmids
Experimental Green Colonies.
LB + AMP Plate: Bacteria + plasmids
Experimental Red Colonies
What was the first animal ever cloned in a lab using embryos? Approximately when?
Frog 1950s
What was the first animal ever cloned from adult tissue? Approximately when?
Frog 1962
What are some applications of stem cells?
Generation of neurons to treat Alzheimer's or Parkinson's Disease. Spinal Cord injuries. Generation of Insulin secreting cells. Used to treat heart disease and stroke. Generate complete organs (i.e. livers, kidneys, eyes, hearts, parts of the brain, etc).
What are the benefits of IVF?
Help infertile couples (or those with difficulty) conceive. Pre-implantation genetic Diagnosiss (PGD) Genetic Modification of human embryos (in the future).
Therapeutic Cloning
Involves growing new tissues or organs.
What is Red Fluorescent Protein? (Where do they come from and why are they useful in biology?)
Is a versatile biological marker for monitoring physiological processes, visualizing protein localization, and detecting transgenic expression in vivo.
What is Gene Therapy?
Is an experimental technique that uses genes to treat or prevent disease.
When/ Where is PGD done?
Is done in a dish (in vitro) a few days after the embryos have developed.
What is Gene Regulation?
Is the informal term used to describe any mechanism used by a cell to increase or decrease the production of specific gene products (protein or RNA). Cells can modify their gene expression patterns to trigger developmental pathways, respond to environmental stimuli, or adapt to new food sources.
What is arabinose's role in our experiment?
It activates transcription of the bacteria to pick up plasmid.
Why is E. coli popular to work with in the lab?
It is easy and inexspensive to grow.
LB + AMP plate: Bacteria - plasmids
Negative Control No Growth
Have humans (adults) been cloned from adult cells in the lab?
No.
What is meant by "resistance to antibiotics"?
Occurs when bacteria change in some way that reduces or eliminates the effectiveness of drugs, chemicals, or other agents designed to cure or prevent infections. The bacteria survive and continue to multiply causing more harm
What is the benefit of performing PGD?
PGD can test for more than 100 different genetic conditions. The procedure is performed before implantation thus allowing the couple to decide if they wish to continue with the pregnancy. The procedure enables couples to pursue biological children who might not have done so otherwise.
LB Plate: Bacteria + plasmids
Positive Control Yellow Lawn
LB Plate: Bacteria - plasmids
Positive Control Yellow Lawn
What is Pre-Implantation Genetic Diagnosis (PGD)?
Refers to the genetic profiling of embryos prior to implantation, and sometimes even of oocytes (eggs) prior to fertilization. Part of IVF. i.e. gender selection, chromosomal number analysis, identification of disease predispositions.
How is Gene Therapy done?
Replacing a mutated gene that causes disease with a healthy copy of the gene. Inactivating, or "knocking out," a mutated gene that is functioning improperly. Introducing a new gene into the body to help fight a disease.
What are Induced Pluripotent Stem (IPS) cells?
Reprogrammed adult cells.
Examples of Tissue Regeneration?
Salamanders can regenerate tissue at sites of injury (i.e. leg cut off, they can regrow it) Planaria have neoblasts that can potentially regenerate the whole organism. Skin cells in humans. Liver is capable of natural regeneration.
What was the process through which Dolly was cloned?
Somatic Cell Nuclear Transfer
What is the inner cell mass?
Source of embryonic stem cells. Source of pluripotent cells.
What is a blastocyst?
Source of pluripotent stem cells.
What are the materials required for PCR and what are their roles?
Taq Polymerase: builds DNA dnTPs: building blocks of DNA MgCl2: required enzymatic co-factor. Primers: control; targets and binds to tubulin gene so it gets copied; says to make a lot of protein.
What is "Germ-Line Gene Therapy"?
The altering of genes in reproductive cells (sperm or egg) in order to affect their function in any offspring that may be created.
What is the role of calcium chloride and the role of the heat shock in bacteria transformation?
The calcium chloride treatment makes cells competent. The heat shock encourages cells to keep any plasmids that they have recently acquired.
What is Genetic Engineering (GE)?
The direct manipulation of an organism's genes by technological intervention/manipulation. Is a phrase used to describe some modern techniques in molecular biology. (i.e. recombinant DNA technology) Tech used to alter the genetic materials of living cells and organisms in order to make them capable of producing new substances or perform new functions. Is a permanent change to the genetic makeup of an organism.
What actually causes the change in phenotype (physical trait) when a cell is transformed?
The introduction of new DNA.
What is IVF?
The mixing of eggs with sperm in a laboratory dish in order to achieve conception.
What is the Process of Transformation? (what does it mean?)
The process of an organism taking up DNA from the environment. The additional DNA isn't necessary for survival of the organism but can lead to a genotype and/or phenotype change. Many plasmids contain genes that are advantageous to the organism.
What is Selective Breeding?
The selection of certain seeds or animals for reproduction in order to influence the traits inherited by the next generation.
What is Epigenetics?
The study of changes in organisms caused by modification of gene expression rather than alteration of the genetic code itself.
Differences between Genetic Engineering and Gene Therapy?
The term "gene therapy" is usually reserved for the process of genetically modifying humans; while modified plants/animals/bacteria/fungi are "genetically engineered." Note: DNA molecules are "genetically engineered" in the laboratory. The process of genetically engineering plants and animals usually takes at the earliest stages of life (embryonic stage); while "gene therapy" is a process of genetically modifying an adult person or child (but long after embryonic stage). Side note: the process of genetically altering human sperm, egg or embryo is usually referred to as "Germ-line Gene Therapy". A genetically engineered organism is "entirely" genetically modified - all cells contain the altered DNA - even if a new trait (or protein) is expressed in one tissue (like human protein made in goat's milk); while "gene therapy" changes the DNA of some cells of an adult person (lung cells, skin cells, heart cells, etc.) making them a chimera.
How are stem cells and tissue regeneration related?
They both renew/repair damaged cells in the body of the organism.
Why are IPS cells an "alternative" to embryonic stem cells?
They de-differentiate (reprogram) adult stem cells.
How do transgenic animals differ from an animal hybrid?
Transgenic animals have specific traits inserted into their genes whereas hybrids are the offspring of two species. i.e. Transgenic mice with cell receptor for polio; Liger- tiger and lion hybrid.
How does US policy towards the sale and labeling of GMO differ from other parts of the world?
U.S: bill passed in August 2016 requiring companies to disclose whether their product contains GM ingredients. EU: any product that contains more than 0.9% of GM ingredients must be labeled; each ingredient must indicate either "genetically modified" or "produced from genetically modified..."
Why is PCR such a useful technique in biotechnology?
Used to amplify trace amounts of DNA, to study genes, and to detect differences in DNA sequences. Used to determine if a person carries a risk for certain genetic diseases. Used to identify an organism or an individual.
What are competent cells?
When an organism has the ability to be transformed, take up DNA from the environment.
Have human embryos been created through somatic cell nuclear transfer?
Yes, in May 2013.
Which of the following statements about CaMV35S is NOT correct? a) Is DNA from bacteria. b) Is a target DNA region in our experiment. c) Is a DNA region known as a promoter. d) Is commonly used in the genetic modification of crops.
a) Is DNA from bacteria.
What is the role of Ampicillin in this experiment? a) It selectively allows only E. coli that picked up plasmid to grow. b) It activates the gene for Green Fluorescent protein in E. coli. c) It is required for the growth of normal E. coli. d) It prevents the growth of genetically transformed E. coli.
a) It selectively allows only E. coli that piked up plasmid to grow.
Reaction occurs in vitro: a) PCR. b) DNA Replication. c) Both.
a) PCR.
If the bacteria died during Heat Shock, you would expect: a) no bacteria on the LB plate for the E. coli + plasmids sample. b) a lawn of bacteria on the LB plate for the E. coli - plasmids sample. c) colonies of bacteria on the LB plate for the E. coli - plasmids sample. d) colonies of bacteria on the LB + Amp plate for the E. coli + plasmids sample.
a) no bacteria on the LB plate for the E. coli + plasmids sample.
If a yellow colony is found on the LB + Amp plate from the E. coli + plasmids sample, the bacteria in the yellow colony contains: a) pGLO. b) pRED. c) both pGLO and pRED. d) no plasmid.
a) pGLO.
What was the purpose of the Tubulin primer in our experiment? a) It will indicate whether or not our food is genetically modified. b) It allows us to amplify the Tubulin gene. c) It acts as a negative control for our PCR reaction. d) B and C only. d) All of the above.
b) It allows us to amplify the Tubulin gene.
Regarding the calcium chloride treatment, if calcium chloride is NOT added to the bacteria: a) the heat shock will kill the bacteria. b) the bacteria will be unable to pick up the plasmid. c) all of the bacteria will be ampicillin resistant. d) the bacteria will be unable to grown on an LB plate.`
b) The bacteria will be unable to pick up the plasmid.
What was the purpose of putting E. coli onto LB plates (with neither Ampicillin nor Arabinose)? a) To show that the E. coli were genetically modified. b) To show that the E. coli were living. c) To induce production of the GFP protein. d) To induce expression of the antibiotic resistance gene.
b) To show that the E. coli were living.
In this experiment, what was the purpose of the bacteria sample that was NOT given any plasmid? a) to determine if transformed bacteria could kill untransformed bacteria. b) as a comparison to prove that the plasmids cause the changes in traits. c) to determine if the plasmid was toxic to the bacteria. d) as a comparison sample to verify that the plasmids allow growth on Luria Broth (LB).
b) as a comparison sample to prove that the plasmids cause the changes in traits.
If the Ampicillin resistance gene on all of the plasmids was mutated and not functioning, you would expect: a) no bacteria on the LB plate for the E. coli - plasmids sample. b) no bacteria on the LB + Amp plate for the E. coli + plasmids sample. c) a lawn of bacteria on the LB + Amp plate for the E. coli + plasmids sample. d) colonies of bacteria on the LB + Amp plate for the E. coli - plasmids sample.
b) no bacteria on the LB + Amp plate for the E. coli + plasmids sample.
What is produced by the Ara C gene on the plasmid pGLO? a) Red Fluorescent protein. b) Green Fluorescent protein. c) A protein that binds to the Arabinose promoter. d) An enzyme that breaks down Ampicillin.
c) A protein that binds to the Arabinose promoter.
What is the role of Arabinose in this experiment? a) Arabinose serves as a negative control. b) Arabinose improves the efficiency of the bacteria to pick up plasmid. c) Arabinose activates transcription of the gene for Green Fluorescent protein. d) Arabinose kills any E. coli that did not pick up plasmid.
c) Arabinose activates transcription of the gene for Green Fluorescent protein.
All of the following statements are true EXCEPT: a) The enzyme RNA polymerase transcribes the RFP gene into an RNA message that gets translated into RFP protein by a ribosome. b) The sugar Arabinose binds to the Ara C protein and changes the protein's shape at the promoter allowing transcription of the GFP gene. c) You attempted to alter both the genotype and phenotype of E. coli. d) Both the GFP and RFP genes are only visible under UV light.
d) Both the GFP and RFP genes are only visible under UV light.
If the ampicillin on an LB + Amp plate is too old and not effective, then: a) no E. coli will grow. b) colonies of E. coli containing pRED will grow. c) colonies of E. coli containing pGLO will grow. d) a lawn of E. coli will grow.
d) a lawn of E. coli will grow.
The gene for RFP on the plasmid, pRED, is regulated by: a) the origin or replication. b) the Ampicillin resistance gene. c) the Arabinose promoter. d) the gene for RFP is not regulated.
d) the gene for RFP is not regulated.
