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General Bio I > Chapter 20-21 Biotechnology and Genomics > Flashcards

Chapter 20-21 Biotechnology and Genomics Flashcards

(103 cards)

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1
Q

What is genetic engineering?

A

extraction and reconfiguration of DNA

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2
Q

What is the goal of genetic engineering?

A

to impact the nucleic acid by recombing DNA

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3
Q

What is recombinant DNA?

A

taking DNA from different locations and putting it into one nucleic acid molecule

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4
Q

What is a natural example of recombinant DNA?

A

crossing over and viruses

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5
Q

Which areas can genetic engineering be applied to?

A

medicine, pharmacy, environmental, forensic analysis, and agriculture

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6
Q

What are restriction enzymes?

A

enzymes floating around recognizing original DNA and destroying foreign DNA

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7
Q

What do restriction enzymes need in order to work?

A

a specific sequence that is from the bacterial cell

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8
Q

What are the sequences that restriction enzymes recognize called?

A

palindromic sequences

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9
Q

What do restriction enzymes do the recognizable DNA?

A

cleaves the DNA to make two strands

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10
Q

After restriction enzymes cleave the DNA, what is created?

A

two strands with sticky ends

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11
Q

What are sticky ends?

A

float around and recombine/reanneal with their complementary strands

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12
Q

What are restriction enzymes used to make for humans?

A

insulin

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13
Q

How do restriction enzymes make insulin?

A

restriction enzymes cut the bacteria DNA, then attach the human insulin gene via a vector on the sticky ends and combine the DNA so bacteria synthesize insulin

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14
Q

What are polymorphisms?

A

Variations in DNA sequence are called

polymorphisms

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15
Q

What are RFLPs

A

restriction fragment length
polymorphisms — a type of polymorphism that results from variation in the DNA sequence recognized by restriction enzymes.

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16
Q

What is gel electrophoresis?

A

separates molecules based on their size by the use of a gel matrix and electric currents

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17
Q

What can gel electrophoresis be used on?

A

DNA, RNA, proteins

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18
Q

How is DNA used in gel electrophoresis?

A

since DNA has a negative charge on the phosphate, they are placed in the holes (aka wells) on one end of the gel and a negative current pushes the DNA to the positive side

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19
Q

What is the gel on gel electrophoresis?

A

agarose (polysaccharide polymer)

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20
Q

How are restriction enzymes and gel electrophoresis related?

A

comparing DNA of organisms requires the same restriction enzymes to cut the same segment of DNA from both organisms to compare at identification points

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21
Q

In the gel electrophoresis, what do different band lengths signify under UV light?

A

shorter DNA moves faster and towards the other end of the gel and longer DNA is more towards the wells

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22
Q

Why is gel electrophoresis important?

A

helps determine relatedness in different species and organisms, DNA fingerprinting

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23
Q

What is Polymerase Chain Reaction (PCR)?

A

helps produce many copies of a

the specific target segment of DNA

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24
Q

What do you need for PCR?

A

the DNA portion to copy, a buffer to put it in, primers, DNA polymerase, DNA nucleotides

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25
Does PCR have to happen in a cell?
NOPE, it can happen in a test tube
26
What kind of DNA polymerase is used for PCR?
heat resistant Taq polymerase
27
Where is the Taq polymerase from?
from bacteria that originally lived at boiling temperatures
28
What is the first step of PCR?
heat up DNA to separate the strands (acts like helicase)
29
What is the second step of PCR?
annealing involves designing primers that specify the target sequence and having those primers hydrogen bond to target at a certain temp
30
What is the third step of PCR?
DNA synthesis --> make more copies of DNA using DNA polymerase using DNA nucleotides at a certain temp
31
Why is PCR important?
DNA fingerprinting and disease diagnosis (covid-19)
32
Single genes are cloned using what?
plasmids in bacterial cells
33
What are plasmids?
small, independent, and circular mobile pieces of DNA that contain nonessential genes
34
Are plasmids a part of the bacterial genome?
no
35
What are vectors?
DNA molecule used as a vehicle to artificially carry foreign genetic material into another cell
36
How does single gene cloning work?
put gene on vector and insert into plasmid then put back into the cell to create more copies of plasmids
37
What is cloning organisms also called?
reproductive cloning
38
What does cloning use?
nuclear transplantation
39
What is nuclear transplantation?
the nucleus of an unfertilized egg cell is replaced with the nucleus of a differentiated cell from an adult organism
40
What can vectors lead to?
GMOs (the first GMO food was a flavor savor tomato)
41
How did cloning animals work? aka Dolly
normal adult nucleus taken from the udder of sheep 1, DNA extracted, egg cell from sheep 2 had sheep DNA put into it and was put into an embryo
42
What are stem cells?
an unspecialized cell that can reproduce indefinitely and can differentiate into specialized cells
43
How do you isolate stem cells?
can be isolated from early embryos at the blastocyst stage (ball that is hollow, stem cells on the inside)
44
Where are stem cells found in adult humans?
bone marrow, but they only form blood related cells
45
For stem cells, what determines what cell type it will become?
transcription factors
46
What is a totipotent stem cell?
can divide into all cell types in an organism
47
What is a pluripotent stem cell?
can divide into most, or all, cell types in an organism, but cannot develop into an entire organism on their own
48
What are IPS cells?
artificial pluripotent cells that have been developed by making a specialized cel become a stem cell again via 4 types of enzymes/viruses
49
What is CRISPR-Cas9?
Clustered Regularly Interspaced Short | Palindromic Repeats
50
What does CRISPR-Cas9 use?
uses short segments of DNA that are palindromic (sequence of letters that read the same from left to right) that has spacer DNA in between the segments
51
What is the spacer DNA in CRISPR-Cas9?
not identical DNA (unique) that matches with viral DNA and cas genes associated with helicase and nucleases
52
What does CRISPR-Cas9 act as?
a tiny immune system
53
When the virus injects their DNA into the bacteria, what happens in CRISPR-Cas9?
CRISPR complex transcribes proteins w/ crRNA/gRNA and the cas proteins break the viral DNA up and embedded it into the CRISPR sequence
54
What does the cas9 protein do?
cuts DNA using CRISPR RNA (gRNA) to inactivate the gene and even mutate it
55
What is the first generation method for genomic sequencing?
Sanger Method
56
What type of nucleotide is used by the sanger method?
dideoxyNTP (ddNTP)
57
What is a ddNTP?
nucleotide that DOESN'T have any oxygen on 3'
58
What does 3' position on a nucleotide determine?
its the backbone to where nucleotides in the sequence can link up to where OH usually is
59
What is ddNTP used for?
termination of synthesis of DNA in the Sager method of DNA sequencing
60
What mechanism is the Sanger method based on?
DNA synthesis/replication
61
What do you need for the Sanger method?
unknown template strand copies of DNA from 3' to 5, primers that are fluorescently labeled, DNA polym, dNTP (large amount), and ddNTP (small amount)
62
What is the Sanger method also called?
chain termination method
63
What are ddNTPs called?
chain terminators
64
What happens in the Sanger method?
separate dNTPs put into 4 test tubes, put ddNTP into it
65
What happens when ddNTP is added to the test tubes?
number fragments of copied DNA that is terminated at certain base pairs (ex: TAAGTCCCT turns to A, TAAGA, TAAGTCCA) according to how many nucleotides are there
66
What is the result of the Sanger method?
DNA fragments with various lengths
67
What happens to the DNA fragments in the Sanger method?
put into gel electrophoresis to separate the lengths of the DNA fragments, read from smallest to largest
68
What is the second generation of DNA sequencing?
NGS (next generation sequencing)
69
What happens in NGS?
short-read sequences from about 50 base pairs to 300 base pairs
70
What is the gold standard for gene sequencing?
sanger method
71
What is the first step in NGS?
DNA gets fragmented and replicated with adaptors on each end of the fragment
72
What is the second step of NGS?
fragments get put onto a glass slide with sequences that are complementary to the adaptors so that the fragments go near the complimentary base pair
73
What is the third step of NGS?
make copies of DNA fragments using polymerase on the slide in the local region and add fluorescent dNTPs
74
What is the fourth step of NGS?
as the sequence is getting synthesized, computer reads fragment as it is getting synthesized and aligns the overlapping fragments to make one genome
75
What is the 3rd generation of genetic sequencing?
nanopore and smrt
76
What happens in nanopore 3rd generation of sequencing?
DNA is fed into an artificial membrane and electric current is sent through the membrane, the nucleotides are disrupted at a specific voltage
77
What happens in SMRT 3rd generation sequencing?
DNA broken into fragments, fluorescent added to the end of phosphate, put into little wells, DNA polymerase lights up the different nucleotides
78
What is genomics?
study of whole sets of genes, structure, function, evolution, and mapping of genomes
79
How much of the DNA is used for the coding sequence?
2 percent
80
What is intergenic DNA?
rest of the DNA that is not coding and is found between genes
81
What are the genetic markers?
single nucleotide polymorphism and short tandem repeats
82
What are SNPs?
one base-pair track which is found in the coding region
83
What are STRs?
little sequence repeats that differ from person to person
84
What are transposable elements?
makeup 45% of the human genome and they are genes that are able to jump to another position on the genome
85
What are the types of transposons?
DNA transposon and retrotransposon
86
What are DNA transposons (eukaryotes)?
transposase cuts and pastes gene from point to point b
87
What are retrotransposons (3 classes)?
use of reverse transcriptase to copy and paste so that genome can increase in size and change profiles
88
What are human's cousins?
chimp that has 24 chromosomes instead of 23 (humans had a fusion)
89
Where do chromosomal changes happen?
crossing over in meiosis for recombination
90
How does divergence happen?
cells deliver different chromosomes to isolate organisms
91
How do changes to chromosomes help speciation?
creates new species that are able to adapt to the environment
92
What are some types of chromosomal arrangements?
can delete or duplicate regions of a chromosome when similar regions of genes are crossed over at the wrong location to create multi family genes
93
What is an example of multifamily genes?
the globin genes because there is alpha and beta
94
What did the globin genes diverge from?
their ancestral globin gene (400-450 mill years ago)
95
When are the globin genes expressed?
during embryonic, fetal, and adult stages of development
96
What are the types of globin genes?
alpha and beta (polypeptides)
97
What are pseudogenes?
genes that don't work anymore and are leftover from evolution due to mutations
98
How are genes with new functions created?
in exon shuffling
99
What happens in exon shuffling?
mistakes in meiosis can duplicate or reposition exons in a more permanent
100
What is an example of exon shuffling?
tpa gene (blood clotting)
101
What are palindromes?
sequences of DNA that are repeated and spaced
102
What do the cas genes have?
helicases and nucleases (that cut DNA)
103
Where do restriction enzymes bind to?
restriction sites (DNA sequence)

General Bio I (18 decks)

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