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BIOC192 > Molecular Biology > Flashcards

Molecular Biology Flashcards

1
Q

Does insulin require PTMs

A

No

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2
Q

A person’s genotype can be determined by

A

PCR-RFLP, fluorescence based methods, sequencing methods

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3
Q

What system is insulin best produced in

A

Bacteria

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4
Q

Does EPO require PTMs

A

Yes, glycosylation

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5
Q

What system is EPO best produced in

A

Eukaryotic system (mammalian) : Chinese hamster ovary cells

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6
Q

Does antithrombin require PTMs

A

Yes (carboxylation)

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7
Q

What system is antithrombin best produced in and why

A

Transgenic animal (goats, pharming). Carboxylation is tricky so won’t occur in a cell culture

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8
Q

What is the central dogma of molecular biology

A

DNA -> mRNA -> protein

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9
Q

What is the basic structure of all genes

A

Promoter region and transcribed region

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10
Q

What is a gene

A

Region of DNA which makes one mRNA molecule

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11
Q

What is different about prokaryotic genes

A

No introns, prokaryotes don’t have the ability to splice. No nucleus, transcribe and translate DNA simultaneously

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12
Q

What is added to mRNA to prevent degradation in eukaryotic cells

A

5’G cap and 3’ poly-A tail

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13
Q

What is present in the promoter region of a gene

A

Transcription factor binding sites and RNA polymerase binding site

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14
Q

What are transcription factors

A

Proteins that bind DNA and control promoter binding of RNA polymerase (can have more than one simultaneously)

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15
Q

What are the two types of transcription factors

A

Activators (turn on), repressors (turn off) by blocking RNA polymerase from binding

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16
Q

How do transcription factors work

A

Use physical connections to turn genes on/off

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17
Q

Why is transcriptional control important

A

So that cells can have different genes turned on for their different functions

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18
Q

Which transcription factors are present differs:

A

Between cells and over time

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19
Q

What are ribosomes made of

A

rRNA and protein: work like enzymes to catalyse protein synthesis (but aren’t actually enzymes)

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20
Q

What is meant by the universal genetic code

A

Codons code for the same amino acids in all organisms

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21
Q

What are the three stages of translation

A

Initiation, elongation, termination

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22
Q

What happens in initiation of translation

A

Ribosome, mRNA and first tRNA come together to form translation initiation complex

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23
Q

What happens in elongation of translation

A

Ribosome moves along mRNA adding amino acids to chain

24
Q

What happens in termination of translation

A

Stop codon is reached, translation complex breaks apart

25
What are the only non-redundant amino acids
Met and Trp
26
What is genetic variation
Differences at the same locus between DNA sequences of members of the same species
27
Why is genetic variation important
Helps us determine who is who, helps a species survive, helps organisms adapt to their environment
28
What is a genetic variant
A specific difference between individuals
29
What are the two types of genetic variants we need to know
SNP and InDel
30
What might a variant in an intron result in
Different expression of the gene (different amount produced) (Stop codon can't be coded for in an intron)
31
What is a missense mutation
One different amino acid coded for
32
What is a nonsense mutation
Early stop codon coded for
33
What do the consequences of a missense mutation depend on
Where in the protein it occurs (e.g active site), how chemically different amino acids are (e.g polar charged, non polar), whether amino acid breaks essential structure (e.g glycine helix breaker, cysteine disulfide bond)
34
What does the HBB E6V variant result in
Glutamic acid to valine, causes aggregation of haemoglobin due to hydrophobic interactions, sickle cell anaemia
35
What is HBB
Haemoglobin B subunit gene
36
What does the HBB H64Y variant result in
Histidine to tyrosine, Fe2+ of haem able to be oxidised to Fe3+, can't bind oxygen
37
The consequences of a missense mutation to the organism depend on
How important the protein's job is, whether another protein can compensate for the lost function, whether the protein lost or gained a function, the inheritance pattern of the protein
38
What mode of inheritance do gain of function alleles have
Dominant
39
What mode of inheritance do loss of function alleles have
Recessive
40
What is retinitis pigmentosa
Retinal degenerative disease caused by variants in rhodopsin receptor protein. Dominant inheritance pattern, one variant allele, gain of function
41
What is phenylketonuria
Monogenic disease resulting in disruption of pathway from phenylalanine to tyrosine, resulting in a build up of phenylalanine causing brain damage
42
What processes enable genotyping
PCR-RFLP, fluorescence based methods, genome sequencing
43
What components are needed in PCR
DNA sample, primers, DNA polymerase (heat tolerant), DNA nucleotides
44
Gain of function alleles often show a ______ inheritance pattern
Dominant
45
Loss of function alleles often show a _______ inheritance pattern
Recessive
46
What type of disease is retinitis pigmentosa
Monogenic gain of function
47
What type of disease is PKU
Monogenic loss of function
48
What are the steps of PCR
Denaturation, annealing, extension
49
What components are needed in PCR
DNA sample, primers, polymerase (heat tolerant DNA polymerase enzyme), nucleotides (dNTPs)
50
What are restriction enzymes
Proteins isolated from bacteria that cut DNA. Each enzyme recognises a palindromic sequence (4-6 bases long)
51
How does gel electrophoresis work
Smaller fragments travel further through gel, toward positive anode
52
What are the common genotyping methods
PCR-RFLP, fluorescence based methods, genome sequencing
53
How does PCR-RFLP work
PCR primers amplify DNA surrounding variant of interest, restriction enzyme cuts specific sequence, fragments move through gel with smallest one going the furthest
54
How do fluorescence based methods work
PCR primers amplify DNA of interest, probes anneal to one of the two alleles, each probe has a different fluorescent dye attached. As polymerase amplifies the DNA the probe is broken down and releases the dye
55
When would you use genome sequencing to genotype an organism
When you don't know where to look in a genome
56
How does genome sequencing work
Determine DNA sequence of whole genome, then zoom in on genetic variants of interest
57
What is gene therapy
Using viral vectors to insert a recombinant gene into patient cells

BIOC192 (5 decks)

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