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Molecular Biology Flashcards

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1
Q

What does northern blotting stain for?

A

RNA

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2
Q

What does western blotting stain for?

A

Proteins

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3
Q

What technique is used to separate DNA?

A

Gel electrophoresis

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4
Q

How are DNA molecules separated, which property?

A

By size

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5
Q

Which gel is better for small fragments of DNA, polyacrilamide or agarose?

A

Polyacrilamide

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6
Q

What is added to DNA to make it detectable?

A

A probe, which is usually radioactive or fluorescent

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7
Q

What method of detection is used for in situ proteins?

A

Specific antigen- antibody interaction

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8
Q

What is a primer?

A

A short single stranded piece of DNA at the starting point

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9
Q

The products of DNA synthesis have different sizes and thus betray the template sequence. Explain how.

A

The normal nucleotides needed for DNA synthesis are spiked with terminal nucleotides
The terminator nucleotides are found in the DNA strand but they prevent the subsequent addition of nucleotides

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10
Q

What type of enzymes are required to fragment DNA?

A

Restriction enzymes

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11
Q

What are the characteristics of restriction enzymes?

A
  1. Each enzyme recognises a specific sequence

2. Restriction enzyme cuts at a specific location in its recognition site

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12
Q

What does PCR stand for?

A

Polymerase chain reaction

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13
Q

What is the key feature of PCR that makes it useful?

A

It is able to reliably amplify DNA sequences from a minute amount

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14
Q

Do we need to know the start and end bases of the sequence of DNA for PCR to work?

A

Yes

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15
Q

Other than DNA templates, what 3 things are required for PCR to work?

A
  1. Primers
  2. A machine that can vary its temperature
    Free nucleotides
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16
Q

What are the 3 stages of PCR?

A
  1. Denaturation
  2. Addition of primers- annealing
  3. Elongation of primers
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17
Q

What feature of the PCR sequence allows us to manipulate DNA?

A

The addition of primers into the DNA strand. This is because mutations in the primers will be built maintained in the new DNA sequence.

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18
Q

What is meant by cloning?

A

Generation of an unlimited supply of a genetic sequence and be able to maintain it outside the its biological context

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19
Q

Why are bacteria so good for cloning?

A
  1. They are easy to grow in large quantities
  2. Easy to purify DNA from them
  3. They have brief life spans
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20
Q

What is a vector? And give an example

A

A vector is an isolated piece of DNA which is maintained and replicated by the host cell. Plasmid.

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21
Q

What sort of genetic code do bacteria contain. Haploid or diploid?

A

Haploid

22
Q

What do bacteria need to grow on?

A

Salts, simple carbohydrates, trace minerals

23
Q

If a bacterium is has a mutation inhibiting a part of the bio synthetic pathway, will the it grow on a minimum medium?

A

No, only on complete medium

24
Q

What is an auxotroph?

A

A bacterium that is unable to synthesise essential nutrients because of mutations

25
Q

Can genetic material be passed between bacteria in both directions?

A

No, only in one direction

26
Q

What is the role of a fertility factor plasmid in bacteria?

A

It is a factor which informs the other bacteria that it is able to conjugate with f recipient bacteria

27
Q

How does fertility factor effect changes in chromosomes?

A

By Hugh frequency recombination

28
Q

Why do organisms allow for gene expression to be regulated?

A

So that they can adapt to changes in the environment and maintain homeostasis

29
Q

What is an Operon?

A

A group of highly packed genes with closely related biochemical functions

30
Q

What conditions are required for lacoperon to be operated and what will the genes be able to do?

A

Lactose needs to be present with no glucose.

Lac Operon will produce proteins which will break down lactose for energy

31
Q

How does lactose cause the lac Operon to function?

A

Lac Operon binds to the lac repressor, this frees up the operator, allowing for cap assisted transcription by RNA polymerase

32
Q

What effect does tryptophan have on the trp Operon

A

It represses it

33
Q

What is a housekeeping gene?

A

These are Genes that are produced in every cell type and code for RNA and proteins that code for universal cell functions

34
Q

What are differentials expressed protein coding genes?

A

These proteins are regulated and only appear in certain cells

35
Q

Selectivity if gene expression is mediated primarily at the level of transcriptional initiation- how?

A
  1. Binding of general transcription factors to the TATA box in the promoter gene sequence
  2. These distort the promoter DNA sequences, allowing the RNA polymerase 2 to bind to the transcription initiation site
36
Q

Why does the GLi 3 transcription factor regulate?

A

Digit numbers in mice and humans- component of the hedgehog signalling pathway

37
Q

What are the initiation codons?

A

AUG (met)

38
Q

What is the termination codes?

A

UAA,UAG,UGA

39
Q

What is the code for tryptophan?

A

UGG

40
Q

Why is the genetic code believed to be a degenerate code?

A

Because most amino acids can be coded for more than one codon

41
Q

What is the role of tRNA?

A

It has anti codons on it which bind to specific codons, to attach the corresponding amino acid to the protein

42
Q

What happens when a new tRNA is added to a ribosome?

A

A hydrogen bond is formed between the amino acids in p and a sites
The E site empties of the redundant tRNA from the triplet code 2 places earlier

43
Q

When a protein folds, what is notable about the structure it forms, with reference to hydrophobicity

A

They hydrophobic side chains are located in the inside of the proteins

44
Q

What 3 forces hold proteins in their secondary and tertiary structures?

A

Van dear waals
Ionic
Hydrogen bonds

45
Q

How many amino acids are present per coil in an alpha helix?

A

7

46
Q

What happens during deamination

A

Cytosine loses an amine, causing the formation of uracil

47
Q

What change in amino acid causes sickle cell anaemia

A

Glutamate to valine

48
Q

What is the main role of the cell cycle?

A

Produce to genetically identical daughter cells

49
Q

What are the 4 phases of the cell cycle?

A
  1. M phase
  2. G1 phase
  3. S phase
  4. G2 phase
50
Q

What happens in s phase?

A

DNA replication

51
Q

Why do cell cycle checkpoints operate through negative signalling?

A

Because the the difference in chemical signalling between 45 and 46 chromosomes is so tiny it would be hard to tell the difference

52
Q

What does southern blotting stain for?

A

DNA

biomedical science (3 decks)

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