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

Chapter 20-21 Biotechnology and Genomics Flashcards

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Biology 101 flashcards
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
Q

Does PCR have to happen in a cell?

A

NOPE, it can happen in a test tube

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26
Q

What kind of DNA polymerase is used for PCR?

A

heat resistant Taq polymerase

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27
Q

Where is the Taq polymerase from?

A

from bacteria that originally lived at boiling temperatures

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28
Q

What is the first step of PCR?

A

heat up DNA to separate the strands (acts like helicase)

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29
Q

What is the second step of PCR?

A

annealing involves designing primers that specify the target sequence and having those primers hydrogen bond to target at a certain temp

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30
Q

What is the third step of PCR?

A

DNA synthesis –> make more copies of DNA using DNA polymerase using DNA nucleotides at a certain temp

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31
Q

Why is PCR important?

A

DNA fingerprinting and disease diagnosis (covid-19)

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32
Q

Single genes are cloned using what?

A

plasmids in bacterial cells

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33
Q

What are plasmids?

A

small, independent, and circular mobile pieces of DNA that contain nonessential genes

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34
Q

Are plasmids a part of the bacterial genome?

A

no

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35
Q

What are vectors?

A

DNA molecule used as a vehicle to artificially carry foreign genetic material into another cell

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36
Q

How does single gene cloning work?

A

put gene on vector and insert into plasmid then put back into the cell to create more copies of plasmids

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37
Q

What is cloning organisms also called?

A

reproductive cloning

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38
Q

What does cloning use?

A

nuclear transplantation

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39
Q

What is nuclear transplantation?

A

the nucleus of an unfertilized egg cell is replaced with the nucleus of a differentiated cell from an adult organism

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40
Q

What can vectors lead to?

A

GMOs (the first GMO food was a flavor savor tomato)

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41
Q

How did cloning animals work? aka Dolly

A

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

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42
Q

What are stem cells?

A

an unspecialized cell that can reproduce indefinitely and can differentiate into specialized cells

43
Q

How do you isolate stem cells?

A

can be isolated from early embryos at the blastocyst stage (ball that is hollow, stem cells on the inside)

44
Q

Where are stem cells found in adult humans?

A

bone marrow, but they only form blood related cells

45
Q

For stem cells, what determines what cell type it will become?

A

transcription factors

46
Q

What is a totipotent stem cell?

A

can divide into all cell types in an organism

47
Q

What is a pluripotent stem cell?

A

can divide into most, or all, cell types in an organism, but cannot develop into an entire organism on their own

48
Q

What are IPS cells?

A

artificial pluripotent cells that have been developed by making a specialized cel become a stem cell again via 4 types of enzymes/viruses

49
Q

What is CRISPR-Cas9?

A

Clustered Regularly Interspaced Short

Palindromic Repeats

50
Q

What does CRISPR-Cas9 use?

A

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
Q

What is the spacer DNA in CRISPR-Cas9?

A

not identical DNA (unique) that matches with viral DNA and cas genes associated with helicase and nucleases

52
Q

What does CRISPR-Cas9 act as?

A

a tiny immune system

53
Q

When the virus injects their DNA into the bacteria, what happens in CRISPR-Cas9?

A

CRISPR complex transcribes proteins w/ crRNA/gRNA and the cas proteins break the viral DNA up and embedded it into the CRISPR sequence

54
Q

What does the cas9 protein do?

A

cuts DNA using CRISPR RNA (gRNA) to inactivate the gene and even mutate it

55
Q

What is the first generation method for genomic sequencing?

A

Sanger Method

56
Q

What type of nucleotide is used by the sanger method?

A

dideoxyNTP (ddNTP)

57
Q

What is a ddNTP?

A

nucleotide that DOESN’T have any oxygen on 3’

58
Q

What does 3’ position on a nucleotide determine?

A

its the backbone to where nucleotides in the sequence can link up to where OH usually is

59
Q

What is ddNTP used for?

A

termination of synthesis of DNA in the Sager method of DNA sequencing

60
Q

What mechanism is the Sanger method based on?

A

DNA synthesis/replication

61
Q

What do you need for the Sanger method?

A

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
Q

What is the Sanger method also called?

A

chain termination method

63
Q

What are ddNTPs called?

A

chain terminators

64
Q

What happens in the Sanger method?

A

separate dNTPs put into 4 test tubes, put ddNTP into it

65
Q

What happens when ddNTP is added to the test tubes?

A

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
Q

What is the result of the Sanger method?

A

DNA fragments with various lengths

67
Q

What happens to the DNA fragments in the Sanger method?

A

put into gel electrophoresis to separate the lengths of the DNA fragments, read from smallest to largest

68
Q

What is the second generation of DNA sequencing?

A

NGS (next generation sequencing)

69
Q

What happens in NGS?

A

short-read sequences from about 50 base pairs to 300 base pairs

70
Q

What is the gold standard for gene sequencing?

A

sanger method

71
Q

What is the first step in NGS?

A

DNA gets fragmented and replicated with adaptors on each end of the fragment

72
Q

What is the second step of NGS?

A

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
Q

What is the third step of NGS?

A

make copies of DNA fragments using polymerase on the slide in the local region and add fluorescent dNTPs

74
Q

What is the fourth step of NGS?

A

as the sequence is getting synthesized, computer reads fragment as it is getting synthesized and aligns the overlapping fragments to make one genome

75
Q

What is the 3rd generation of genetic sequencing?

A

nanopore and smrt

76
Q

What happens in nanopore 3rd generation of sequencing?

A

DNA is fed into an artificial membrane and electric current is sent through the membrane, the nucleotides are disrupted at a specific voltage

77
Q

What happens in SMRT 3rd generation sequencing?

A

DNA broken into fragments, fluorescent added to the end of phosphate, put into little wells, DNA polymerase lights up the different nucleotides

78
Q

What is genomics?

A

study of whole sets of genes, structure, function, evolution, and mapping of genomes

79
Q

How much of the DNA is used for the coding sequence?

A

2 percent

80
Q

What is intergenic DNA?

A

rest of the DNA that is not coding and is found between genes

81
Q

What are the genetic markers?

A

single nucleotide polymorphism and short tandem repeats

82
Q

What are SNPs?

A

one base-pair track which is found in the coding region

83
Q

What are STRs?

A

little sequence repeats that differ from person to person

84
Q

What are transposable elements?

A

makeup 45% of the human genome and they are genes that are able to jump to another position on the genome

85
Q

What are the types of transposons?

A

DNA transposon and retrotransposon

86
Q

What are DNA transposons (eukaryotes)?

A

transposase cuts and pastes gene from point to point b

87
Q

What are retrotransposons (3 classes)?

A

use of reverse transcriptase to copy and paste so that genome can increase in size and change profiles

88
Q

What are human’s cousins?

A

chimp that has 24 chromosomes instead of 23 (humans had a fusion)

89
Q

Where do chromosomal changes happen?

A

crossing over in meiosis for recombination

90
Q

How does divergence happen?

A

cells deliver different chromosomes to isolate organisms

91
Q

How do changes to chromosomes help speciation?

A

creates new species that are able to adapt to the environment

92
Q

What are some types of chromosomal arrangements?

A

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
Q

What is an example of multifamily genes?

A

the globin genes because there is alpha and beta

94
Q

What did the globin genes diverge from?

A

their ancestral globin gene (400-450 mill years ago)

95
Q

When are the globin genes expressed?

A

during embryonic, fetal, and adult stages of development

96
Q

What are the types of globin genes?

A

alpha and beta (polypeptides)

97
Q

What are pseudogenes?

A

genes that don’t work anymore and are leftover from evolution due to mutations

98
Q

How are genes with new functions created?

A

in exon shuffling

99
Q

What happens in exon shuffling?

A

mistakes in meiosis can duplicate or reposition exons in a more permanent

100
Q

What is an example of exon shuffling?

A

tpa gene (blood clotting)

101
Q

What are palindromes?

A

sequences of DNA that are repeated and spaced

102
Q

What do the cas genes have?

A

helicases and nucleases (that cut DNA)

103
Q

Where do restriction enzymes bind to?

A

restriction sites (DNA sequence)

General Bio I (18 decks)

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