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Ch 20 Flashcards

1
Q

Genome

A

All genes of a cell, tissue, or organism. The human genome has approximately 25,000 genes.

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2
Q

Host organisms

A

Host organisms are used to grow and replicate the DNA introduced by vectors.
They are often bacteria, but do not have to be.

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3
Q

Denature, Anneal, and Extension are the steps of what process?

A

PCR

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4
Q

Role of Restriction Enzymes

A

Restriction enzymes cleave DNA into smaller segments of various sizes.

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5
Q

(Circle with two different colors) What does the diagram show?

A

Illustration of how DNA fragments and vectors (plasmids) are combined to form recombinant DNA.

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6
Q

Uses of microorganisms in biotechnology

A

To make wine and cheese and other products of fermentation, clean up oil spills or sewage treatment, production of antibiotics/ medicines, selective breeding of livestock, paternity testing, criminal law, GM foods, etc.

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7
Q

Gel electrophoresis

A

Gel electrophoresis is a method for separating DNA fragments based on size.

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8
Q

Step 1 of PCR

A

Heat to separate DNA strands at 95°C. High temperature breaks hydrogen bonds between DNA strands.

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9
Q

Practical goal of biotech

A

Improvement of human health and food production

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10
Q

Movement of DNA Fragments in gel electrophoresis

A

When an electric current is passed through the chamber, DNA fragments move toward the positively-charged electrode.
Smaller fragments travel much faster.

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11
Q

Role of primers in PCR

A

Primers bracket the area to be copied to get a specific sequence

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12
Q

Bacterial Plasmids

A

Small, circular DNA molecules that replicate within bacterial cells; different from the chromosome.

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13
Q

Speed of DNA Segments in gel electrophoresis

A

Smaller DNA segments move faster and farther than larger DNA segments.

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14
Q

What is a DNA ladder?

A

A DNA ladder consists of fragments of known length that allow you to estimate the length of sample fragments.
Reference for estimating DNA fragment sizes.

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15
Q

DNA technology has launched a revolution in the area of:

A

Biotechnology

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16
Q

Genome size

A

Total amount of DNA, measured in the number of base pairs. It is not the same as the number of genes.

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17
Q

Step 2 of PCR

A

Cool to 55°C to allow primers to bind (anneal).

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18
Q

Step 3 of PCR

A

Heat to 72°C so that DNA polymerase extends the 3’ end of each primer.

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19
Q

Sticky ends of restriction fragments are used for what purpose?

A

This process uses complementary base pairing from the sticky ends to create recombinant DNA.

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20
Q

PCR

A

PCR (Polymerase Chain Reaction) is a technique used to amplify small segments of DNA.

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21
Q

Restriction enzymes

A

Restriction enzymes cut DNA at specific sequences, allowing for manipulation of DNA fragments.

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22
Q

How many copies of DNA would you have after Cycle 20 of PCR

A

2^20 = 1,048,576 copies

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23
Q

Function of restriction enzymes in nature

A

These enzymes protect the bacterial cell from viruses by cutting up their foreign DNA.

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24
Q

Definition of Biotechnology

A

The use of living organisms or their components to do practical tasks

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25
DNA vector
A DNA vector is a vehicle used to transfer genetic material to a target cell. Bacterial plasmids and viruses make good vectors.
26
Why are bacteria commonly used as Host organisms in genetic engineering?
Bacteria are commonly used as hosts because DNA can be easily isolated and reintroduced, and bacterial cultures grow quickly, replicating foreign genes.
27
Starting materials for PCR
DNA to be copied, Nucleotides (nucleoside triphosphates), Primers, Taq polymerase
28
Vectors
Carriers for moving DNA from test tubes back into cells.
29
Unions of different DNA sources can be made permanent by adding what enzyme?.
DNA ligase forms covalent bonds between nucleotides.
30
1995: sequencing of the first complete genome; what was it?
The first complete genome of a bacterium was sequenced in 1995.
31
Gel Electrophoresis
Uses electricity to separate DNA fragments or other molecules by size, shape, and/or charge.
32
Analogy for symmetrical sequences
An example of a symmetrical sequence is the word "KAYAK," which reads the same forwards and backwards.
33
How many copies of DNA would you have after Cycle 5 of PCR?
2^5 = 32 copies
34
Each PCR reaction __________ the amount of DNA.
Doubles; A standard PCR sequence of 30 cycles creates over 1 billion copies.
35
Recombinant DNA
DNA from two sources, often two species, combined in vitro into the same DNA molecule.
36
Step 4 of PCR
Repeat steps 1-3 many times.
37
Function of Taq polymerase
Elongate DNA off the 3' end of primers; DNA polymerase isolated from bacteria in hot springs; enzymes can withstand high temperatures of PCR machine
38
Purpose of Polymerase Chain Reaction (PCR)
Allows any piece of DNA to be quickly amplified in vitro.
39
Symmetrical restriction sequences
Most restriction sequences are symmetrical, with the same sequence of 4-8 nucleotides found on both strands but running in opposite directions.
40
Function of restriction enzymes in biotech
Restriction enzymes in biotechnology cut phosphodiester bonds of both strands in a staggered manner, producing single-stranded "sticky ends."
41
sequencing of the Human Genome completed in _____?
The Human Genome Project was completed in 2003, mapping the entire human genetic code.
42
Genetic engineering
The direct manipulation of genes for practical purposes.
43
Why does Each person have two sets of every STR repeat?
Each person has two sets of repeats because they inherit one set from each parent.
44
Describe the need to stain DNA in gel electrophoresis
DNA has no color; a chemical is needed to bind to the DNA fragments. Examples include Sybr safe, GelRed, GelGreen, Ethidium Bromide (EtBr).
45
Why can STRs be used to create a DNA Fingerprint
Short Tandem Repeats (STRs) are used because different numbers of repeats result in fragments of different lengths after PCR that would be seen when run through a gel.
46
How is Viral RNA converted to DNA for PCR?
Viral RNA in the sample is converted to DNA using Reverse Transcriptase enzyme through the process of reverse transcription..
47
What are Short Tandem Repeats (STRs)
STRs are sequences of 2-6 nucleotides that repeat in chromosomes, used in DNA fingerprinting.
48
DNA Fingerprinting: Key Processes
Involves PCR (Polymerase Chain Reaction), restriction enzymes, and gel electrophoresis.
49
Describe the step in human gene cloning for culturing bacteria.
Give bacteria plenty of food and antibiotics. Allow transformed bacteria (would have the gene of interest and gene for antibiotic resistance) to grow, divide, and produce the desired protein (e.g., human insulin).
50
How is it possible that Bacteria can produce insulin through recombinant DNA technology?
The genetic code is universal!
51
Steps to get a DNA fingerprint
1) obtain a DNA sample from sources such as cheek cells, blood, tissue, or semen 2) Use PCR to amplify noncoding regions of DNA where differences are known to occur, such as STRs 3) load DNA samples into wells of gel electrophoresis; must apply stain/probe 4) Use UV light to see the DNA bands/fragments
52
DNA Technologies: DNA fingerprinting (profiling)
DNA fingerprinting is a technique used to identify individuals by characteristics of their DNA.
53
How does gel electrophoresis work?
DNA is loaded at the top in a well with a negative charge. The negative charge allows DNA to move towards the positive electrode during electrophoresis.
54
What should be mixed with agarose for transformation?
Mix in an antibiotic that matches the inserted antibiotic resistance gene.
55
In gel electrophoresis, what do brighter bands indicate?
Brighter bands indicate more fragments of that size are present. This can be used to estimate the concentration of DNA in the sample.
56
In Step 1 in Cloning a Human Gene, explain the steps to isolate the human gene to clone.
Isolate the human gene to clone, such as insulin. Use mRNA from eukaryotic cells (introns removed) as a template to form double-stranded DNA. Process is called reverse transcription and uses the enzyme reverse transcriptase.
57
DNA Fingerprinting uses
Used for crime scenes, paternity testing, and diagnosing diseases.
58
How can a PCR machine can be used to detect viruses?
PCR can detect viruses by converting viral RNA to DNA, using primers to bracket viral DNA, and attaching fluorescent markers to detect fluorescence levels. If it glows, the virus is present (ex: Covid or HIV).
59
In gel electrophoresis, why is DNA mainly separated due to size?
DNA fragments are separated by size because DNA is negatively charged and linear.
60
Describe the transformation step when Cloning a Human Gene
Mix recombinant plasmids with bacteria. Some bacteria will take in the plasmid, becoming transformed.
61
Role of Antibiotic Resistance Gene
The antibiotic resistance gene produces a protein that breaks down the antibiotic.
62
Describe the creation of recombinant DNA for use in Cloning a Human Gene
Insert the gene of interest into a plasmid vector with matching sticky ends and a gene for antibiotic resistance. Use matching restriction enzymes for the gene and plasmid, and add ligase.
63
DNA Technologies: CRISPR
CRISPR is a technology used for editing genomes, allowing researchers to alter DNA sequences and modify gene function.
64
DNA Technologies: Gene Knockout
Gene knockout involves deactivating a gene to study its function by observing the effects of its absence.
65
DNA Technologies: Cloning
Cloning is a process of creating genetically identical copies of biological entities.
66
Describe the protein purification step of gene cloning.
The final step in cloning a human gene for insulin involves lysing/ breaking open the bacteria to extract and purify the protein (ex: insulin) for medical use.
67
Environmental uses of DNA Technology
Microorganisms engineered to break down sewage and oil spills
68
Library of Knockout Organisms
A collection of knockout organisms available for some species, usually mice, to use as a model in research.
69
Applications of DNA Technology in Agriculture
DNA technology is used in agriculture to enhance livestock and plants, often referred to as "Pharm" animals and plants.
70
CRISPR: a genetic engineering tool
CRISPR is a tool that uses a CRISPR sequence of DNA and its associated protein to edit the base pairs of a gene.
71
Functions of CRISPR
CRISPR can correct a gene or knockout a gene.
72
How does the image diagram gene knockout?
Illustrates the difference between a wildtype mouse with functional gene expression and a knockout mouse with no gene expression.
73
Purpose of Gene Knockout
To study the impact of removing a gene from an organism, allowing scientists to learn about that gene's function.
74
Practical Applications of DNA Technology in Medicine
Includes pharmaceutical products like insulin, human growth hormone, and TPA (which dissolves blood clots), as well as disease diagnosis.
75
Diagram showing CRISPR mechanism
The diagram illustrates how CRISPR uses guide RNA and Cas9 to match genomic sequences and edit DNA by deleting or inserting genes.
76
Use of bGH in Livestock
bGH (bovine growth hormone) is used to enhance milk production in livestock.
77
CRISPR's rise in use
CRISPR has become more common since 2012.
78
Growth Hormone in Salmon
Salmon are genetically modified to grow larger due to a modified growth hormone.
79
Genetic Modification in Cattle
Genes are modified to cause the development of larger muscles in cattle.
80
Applications of DNA Technology in forensics
Forensic uses: DNA fingerprints for paternity and criminal cases
81
Gene Knockout
The use of genetic engineering to inactivate or remove one or more specific genes from an organism.
82
Genetic Engineering in Crops
Crops can be genetically modified to be resistant to herbicides and insecticides, ripen or spoil slower, and have enhanced nutritional qualities such as more vitamins
83
CRISPR in medical trials
CRISPR has been used successfully in trials to turn fetal hemoglobin gene back on in a sickle cell patient.
84
what does transgenic mean?
an organism that had another species' DNA put into it
85
Another name for GMO
Transgenic

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