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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
Q

What are the only non-redundant amino acids

A

Met and Trp

26
Q

What is genetic variation

A

Differences at the same locus between DNA sequences of members of the same species

27
Q

Why is genetic variation important

A

Helps us determine who is who, helps a species survive, helps organisms adapt to their environment

28
Q

What is a genetic variant

A

A specific difference between individuals

29
Q

What are the two types of genetic variants we need to know

A

SNP and InDel

30
Q

What might a variant in an intron result in

A

Different expression of the gene (different amount produced)
(Stop codon can’t be coded for in an intron)

31
Q

What is a missense mutation

A

One different amino acid coded for

32
Q

What is a nonsense mutation

A

Early stop codon coded for

33
Q

What do the consequences of a missense mutation depend on

A

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
Q

What does the HBB E6V variant result in

A

Glutamic acid to valine, causes aggregation of haemoglobin due to hydrophobic interactions, sickle cell anaemia

35
Q

What is HBB

A

Haemoglobin B subunit gene

36
Q

What does the HBB H64Y variant result in

A

Histidine to tyrosine, Fe2+ of haem able to be oxidised to Fe3+, can’t bind oxygen

37
Q

The consequences of a missense mutation to the organism depend on

A

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
Q

What mode of inheritance do gain of function alleles have

A

Dominant

39
Q

What mode of inheritance do loss of function alleles have

A

Recessive

40
Q

What is retinitis pigmentosa

A

Retinal degenerative disease caused by variants in rhodopsin receptor protein. Dominant inheritance pattern, one variant allele, gain of function

41
Q

What is phenylketonuria

A

Monogenic disease resulting in disruption of pathway from phenylalanine to tyrosine, resulting in a build up of phenylalanine causing brain damage

42
Q

What processes enable genotyping

A

PCR-RFLP, fluorescence based methods, genome sequencing

43
Q

What components are needed in PCR

A

DNA sample, primers, DNA polymerase (heat tolerant), DNA nucleotides

44
Q

Gain of function alleles often show a ______ inheritance pattern

A

Dominant

45
Q

Loss of function alleles often show a _______ inheritance pattern

A

Recessive

46
Q

What type of disease is retinitis pigmentosa

A

Monogenic gain of function

47
Q

What type of disease is PKU

A

Monogenic loss of function

48
Q

What are the steps of PCR

A

Denaturation, annealing, extension

49
Q

What components are needed in PCR

A

DNA sample, primers, polymerase (heat tolerant DNA polymerase enzyme), nucleotides (dNTPs)

50
Q

What are restriction enzymes

A

Proteins isolated from bacteria that cut DNA. Each enzyme recognises a palindromic sequence (4-6 bases long)

51
Q

How does gel electrophoresis work

A

Smaller fragments travel further through gel, toward positive anode

52
Q

What are the common genotyping methods

A

PCR-RFLP, fluorescence based methods, genome sequencing

53
Q

How does PCR-RFLP work

A

PCR primers amplify DNA surrounding variant of interest, restriction enzyme cuts specific sequence, fragments move through gel with smallest one going the furthest

54
Q

How do fluorescence based methods work

A

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
Q

When would you use genome sequencing to genotype an organism

A

When you don’t know where to look in a genome

56
Q

How does genome sequencing work

A

Determine DNA sequence of whole genome, then zoom in on genetic variants of interest

57
Q

What is gene therapy

A

Using viral vectors to insert a recombinant gene into patient cells

BIOC192 (5 decks)

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