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BIOC 3622 - Biochemistry Lab > Genetically Modified Organisms and PCR > Flashcards

Genetically Modified Organisms and PCR Flashcards

1
Q

What is the name of today’s lab?

A

Genetically Modified Organisms and PCR

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2
Q

What is the acronym PCR?

A

Polymerase Chain Reaction

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3
Q

What is PCR?

A

DNA amplification

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4
Q

Traits possible through genetic modification include herbicidal resistance, ________, fungal resistance, and even increased _______ tolerance.

A

delayed ripening; drought

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5
Q

The ability to genetically modify crops relies on our ability as researchers to both identify and fully understand __________ with the potential to improve our target organism.

A

candidate gene

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6
Q

Using crop plants as an example, it is first important to identify and _____ (clone) for a protein that may bestow beneficial characteristic to the plant.

A

isolate

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7
Q

While the gene’s coding region may only consist of a few _____ base pairs, the inclusion of non-coding regions (introns) can extend the gene’s length well into the range of _______ or more base pairs.

A

hundred; ten thousand

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8
Q

The cloning of an entire gene can be extremely ______, and can take several ____ to complete.

A

laborious; years

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9
Q

Once a gene is successfully ____ for a ___ organism, it must be successfully _____ so that it expresses properly as part of its adopted organism’s _____.

A

cloned; host; engineered; genome

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10
Q

Given that the signals regulating expression of genes may differ greatly between the host and target organism, it is optional that the streamlined in such a way that it will properly manufacture its product regardless of its location. True or False?

A

False. It is imperative that the gene is streamlined.

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11
Q

Given that the signals regulating expression of genes may differ greatly between the host and target organism, it is imperative that the streamlined in such a way that it will properly manufacture its product regardless of its location. True or False?

A

True

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12
Q

This is achieved first by removing any _____ sections from the gene, known as ______, and replacing the gene’s native _________ and ______ sequence.

A

non-coding; introns; promoter; terminator

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13
Q

In modified genes, the promoter and terminator are specific sequences of the gene that serve as the regulatory mechanisms. True or False? If false, how would you make the statement true?

A

False. In *unmodified* genes…

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14
Q

In unmodified genes, the promoter and terminator are general sequences of the gene that serve as the regulatory mechanisms. True or False? If false, how would you make statement true?

A

False. The promoter and terminator are *specific* sequences of the gene.

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15
Q

In unmodified genes, the promoter and terminator are specific sequences of the gene that serve as the regulatory mechanisms. True or False? If false, how would you make the statement true?

A

True

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16
Q

The promoter region serves as a docking station for __________ and acts as a signal for where _________ should begin.

A

RNA polymerase; gene transcription

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17
Q

The terminator serves as a signal to _____ transcription.

A

stop

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18
Q

For genes involved in genetic modification, the natural promoters and terminators may behave unreliably within their new hosts. True or False? If false, how would you make the statement true?

A

True

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19
Q

To circumvent the problem of unreliability among the promoters and terminators, scientists can engineer the construct for a ___________ so that the foreign gene is continually transcribed, and the protein product is produced throughout the entire plant.

A

genetically modified organism (GMO)

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20
Q

For GM crops, the most common promoter is the 35S promoter from the __________ virus (CaMV 35S).

A

cauliflower mosaic

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21
Q

The most common terminator in GM crops is the _________ (___) terminator from Agrobacterium tumefaciens.

A

nopaline synthase; NOS

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22
Q

____ is the process in which a gene’s DNA sequence is copied (______) to make an RNA molecule.

A

Transcription; transcribed

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23
Q

______ is the main transcription enzyme.

A

RNA polymerase

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24
Q

Transcription begins when RNA polymerase binds to a ______ sequence near the beginning of a gene (directly or through helper proteins).

A

promoter

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25
RNA polymerase uses one of the DNA strands (the \_\_\_\_\_\_\_\_\_) as a template to make a new, complementary RNA molecule.
template strand
26
Transcription ends in a process called \_\_\_\_\_\_\_\_\_.
termination
27
A second method is \_\_\_\_\_\_, in which an electric current creates pores within the cellular membranes to allow entry of the engineered DNA.
electroporation
28
A third method involves the use of a ________ that physically shoots gold particles coated with engineered DNA into the plant's cells.
"gene gun"
29
Unfortunately methods 1-3 lack \_\_\_\_\_, and it is necessary to ____ and ____ cells that have been successfully transformed from those that have not.
efficiency; identify; isolate
30
To assist in this process, selectable markers are often inserted in addition to the engineered gene. These markers may include ______ resistance, or visual markers like the gene \_\_\_\_\_\_\_.
antibiotic; Green Fluorescent Protein
31
To properly identify whether or not a plant has been genetically modified, we will be using the ________________ (\_\_\_).
polymerase chain reaction; PCR
32
The PCR reaction can identify ______ of ___ that have been inserted in the GM plant.
sequences; DNA
33
Because genetic engineers only use a small number of regulatory sequences (\_\_\_\_\_\_ and \_\_\_\_\_) to control the expression of inserted genes, _____ can be used to identify a large portion of all GM crops relatively easy.
promoters; terminators; PCR
34
Two of the most common regulatory sequences are the _________ of cauliflower mosaic virus and the _________ of the Agrobacterium tumefaciens.
35S promoter; nopaline synthase terminator
35
What is step 1 to genetically modify an organism?
isolate plant cells
36
What is step 6 to genetically modify an organism?
Grow transgenic plant
37
What is step 6 to genetically modify an organism?
Grow transgenic plant
38
What is step 5 to genetically modify an organism?
Redifferentiate callus
39
What is step 2 to genetically modify an organism?
Grow undifferentiated cells
40
What is step 4 to genetically modify an organism?
Select cells
41
What is step 3 to genetically modify an organism?
Transform callus
42
Which two methods are used to identify which foods and crops contain GM genomes?
ELISA and PCR
43
What is the acronym ELISA?
enzyme-linked immunosorbent assay
44
What is the acronym PCR?
polymerase chain reaction
45
Which method identifies proteins?
ELISA
46
Which test identifies sequences of DNA?
PCR
47
ELISA can test both fresh and old produce. True or false? If false, make the statement true.
False. It can only test fresh produce due to protein degradation during food processing.
48
ELISA can test only fresh produce. True or false? If false, make the statement true.
True
49
ELISA can test only old produce. True or false? If false, make the statement true.
False. It can only test fresh produce.
50
ELISA is accurate yet expensive and can be performed in the field with little expertise. True or false? If false, make statement true.
False. ELISA is inexpensive.
51
ELISA is accurate yet inexpensive and can only be performed in the field with someone that has a lot of expertise. True or false? If false, make statement true.
False. It can be performed in the field with little expertise.
52
ELISA is accurate yet inexpensive and can be performed in the field with little expertise. True or false? If false, make statement true.
True
53
In contrast to proteins, DNA is a relatively ____ molecule, thus DNA fragments can be isolated from highly processed foods and are sufficiently _____ to be ____ by PCR.
stable; intact; amplified
54
In contrast to an ELISA test that is specific to a single crop, a single ____ test can detect ____ of all GM crops.
PCR; 85%
55
Why can a PCR test detect 85% of all GM crops?
Only a small # of regulatory sequences (promoter and terminator) control the expression of the inserted genes, so these sequences are common to the majority of GM crops.
56
When detecting GM foods by PCR, ______ is extracted from test foods, and then two __________ are performed on each test food genomic DNA sample.
genomic DNA; PCR reactions
57
One PCR reaction uses _____ specific to a common plant gene (\_\_\_\_) to verify that viable DNA was successfully extracted from the food.
primers; plant primers
58
No matter whether the food is GM or not, this PCR reaction should sometimes amplify the DNA. True or false? If false, make statement true.
False. PCR reaction should \*always\* amplify the DNA.
59
No matter whether the food is GM or not, this PCR reaction should always amplify the DNA. True or false? If false, make statement true.
True
60
This PCR reaction will only amplify DNA if the test food is \_\_\_\_\_.
GM
61
If the test food is non-GM, then the GMO primers will not be ________ to any within the test food genomic DNA and will not \_\_\_\_\_, so no DNA will be \_\_\_\_.
complimentary; anneal; amplified
62
To find out whether DNA has been amplified or not, the PCR products are ______ on a gel and stained to visualize _____ as bands.
electrophoresed; DNA
63
A ________ ruler is ________ with the samples to allow the sizes of the DNA to be \_\_\_\_.
molecular weight; electrophoresed; determined
64
When detecting GM foods by PCR, ________ is extracted from test foods, and then two ______ are performed on each test food genomic _______ sample.
genomic DNA; PCR reactions; DNA
65
One PCR reaction uses ______ specific to a common plant gene (\_\_\_\_\_\_\_) to verify that viable DNA was successfully extracted from the food.
primers; plant primers
66
No matter whether the food is GM or not, this PCR reaction should sometimes amplify DNA. True or false? If false, make statement true.
False. PCR reaction should \*always\* amplify DNA.
67
No matter whether the food is GM or not, this PCR reaction should always reduce DNA. True or false? If false, make statement true.
False. PCR reaction should always \*amplify\* DNA.
68
No matter whether the food is GM or not, this PCR reaction should always amplify RNA. True or false? If false, make statement true.
False. The PCR reaction should always amplify \*DNA\*.
69
No matter whether the food is GM or not, this PCR reaction should always amplify DNA. True or false? If false, make statement true.
True
70
The other PCR reaction uses _____ specific to sequences commonly found in GM crops (GM \_\_\_\_\_).
primers; primers
71
This PCR reaction will only amplify DNA if the test food is \_\_\_.
GM
72
If the test food is non-GM, then the GMO primers will not be ____ to any sequence within the test food genomic DNA and will not \_\_\_\_, so no DNA will be \_\_\_\_.
complementary; anneal; amplified
73
To find out whether DNA has been ____ or not, the PCR products are _____ on a gel and ____ to visualize DNA as bands.
amplified; electrophoresed; stained
74
A ________ is electrophoresed with the samples to allow the sizes of the ____ bands to be determined.
molecular weight ruler; DNA
75
In today's lab, we see many of the times GM crop and DNA GM... What is the acronym "GM" stand for?
Genetically modified
76
In today's lab, it discusses something called a "callus". What does that mean?
A clump of undifferentiated cells.
77
How do callus cells develop roots, shoot and eventually grow into entire plants?
Growth hormones can be added to the media which trigger the callus to grow.
78
What is 9?
Quantify GM content by real-time
79
What is 8?
Food contains no GMO.
80
What is 3?
Extract DNA
81
What is 1?
Detect GMOs by PCR
82
What is 5?
not viable
83
What is 7?
Food contains GMO
84
What is 4?
viable
85
What is 6?
No conclusions can be made. Repeat test.
86
What is 2?
Detect GMOs by ELISA
87
What are 1 & 2 respectively?
Non-GM food; GM food
88
What are 9 & 10 respectively?
455 bp; No GMO
89
What are 11 & 12 respectively?
455 bp; 200 bp
90
What are 3 & 4 respectively?
Extract genomic DNA; extract genomic DNA
91
What are 7 & 8 respectively?
Plant primers; GMO primers
92
What are 5 & 6 respectively?
Plant primers; GMO primers
93
In this section of the lab, you will ___ DNA from a control ____ food and a grocery store food item, and you will test for the presence of \_\_\_\_\_.
extract; non-GMO; GMOs
94
What are the two most commonly modified food items?
corn and soy based
95
Which control will you process first?
non-GMO
96
During the extraction of the food samples, you will first weigh your food sample, then ____ it with water to make a \_\_\_\_.
grind; slurry
97
You will then add a tiny amount of the slurry to a _____ tube containing _______ matrix and heat it for ___ minutes.
screwcap; InstaGene; 5
98
What are the name of the enzymes that you are releasing from the ground-up sample?
DNases
99
What can the enzyme DNases do to DNA?
degrade the DNA attempting to be extracted
100
The InstaGene matrix is made of ___ charged microscopic beads that _____ or grab metal ions out of solution.
negatively; "chelate"
101
What metal ions are among being chelated?
Mg2+
102
Why is Mg2+ required?
It is a cofactor in enzymatic reactions
103
When DNA is released from your sample without the presence of the InstaGene matrix, the charged beads grab the Mg2+ and make it unavailable to the enzymes that would degrade the DNA you are trying to extract. True or false? If false, make statement true.
False. This only happens if the DNA is released from your sample \*in\* the presence of the InstaGene matrix.
104
When DNA is released from your sample in the presence of the InstaGene matrix, the charged beads grab the Mg2+ and make it available to the enzymes that would degrade the DNA you are trying to extract. True or false? If false, make statement true.
False. The charged beads grab the Mg2+ and make it \*unavailable\* to the enzyme...
105
When DNA is released from your sample in the presence of the InstaGene matrix, the charged beads avoid the Mg2+ and make it unavailable to the enzymes that would degrade the DNA you are trying to extract. True or false? If false, make statement true.
False. The charged beads \*grab\* the the Mg2+.
106
What allows you to extract DNA without degradation?
When DNA is released in the presence of the InstaGene matrix.
107
Boiling the samples _______ these enzymes.
destroys
108
After you _____ the samples to remove the INstaGene matrix and debris, the _____ will contain the intact extracted \_\_\_\_.
centrifuge; supernatant; DNA
109
What will be used as your target DNA?
the extracted DNA
110
When DNA is released from your sample in the presence of the InstaGene matrix, the charged beads grab the Mg2+ and make it unavailable to the enzymes that would degrade the DNA you are trying to extract. True or false? If false, make statement true.
True
111
ALWAYS process the ______ control before the test sample to reduce the risk of \_\_\_\_\_.
non-GMP; contamination
112
PCR has 3 steps. Write them in the correct order.
Denature DNA. Anneal them. Extend them.
113
The most commonly used thermostable DNA polymerase is the TAQ DNA polymerase. True or false? If false, make statement true.
True

BIOC 3622 - Biochemistry Lab (8 decks)

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