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My Packs: UWO (BIO 1002) > Cycle 2 BMP Workshop > Flashcards

Cycle 2 BMP Workshop Flashcards

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Biology 101 flashcards
1
Q

Define:

Gene expression

A

Process when a gene is turned into a function

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2
Q

How does a gene become an RNA transcript?

A

Transcription

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3
Q

How does an RNA transcript become a protein?

A

Translation

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4
Q

Describe:

Northern Blot Technique

A

Tells us if a gene is transcribed (mRNA)

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5
Q

Explain:

The acronym SNOW DROP

A

Southern Blot - DNA
Northern Blot - RNA
OOOOOOOO - OOO
Western Blot - Protein

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6
Q

Describe:

Steps of Northern Blot

A
  1. Extract mRNA
  2. Gel Electrophoresis (separate mRNA strands by size)
  3. Transfer gel to a membrane
  4. Probe the membrane
  5. Visualize the probes
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7
Q

True or False:

In Gel Electrophoresis, the smaller mRNAs will move to negative end faster

A

False, the smaller mRnas will move to positive end faster/further than the larger mRNAs

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8
Q

Describe:

Step 3 in Northern Blot

A

Step 3: Transfer gel to a membrane (since it’s hard to see all mRNA on the gel as it is so thick)
* Place gel in salt solution
* Wick all the mRNAs from gel
* Get onto membrane

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9
Q

Describe:

Step 4 of Northern Blot

A

Step 4: Probe the membrane
* Use single-stranded DNA probes with fluorescent radioactive label (denatured to become single strand)
* Mix DNA probes with the membrane
* ssDNA probes will complementary base-pair with GAL mRNA

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10
Q

How is the fluorescent/radiactive label transferred in Northern Blot?

A

By using an x-ray film

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11
Q

True or False:

Darker lines in Northern Blot means lots of transcript abundance

A

True

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12
Q

Define:

Gene

A

A DNA sequence that codes for an RNA strand

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13
Q

True or False:

All genes code for mRNA

A

False, also codes for rRNA and tRNA

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14
Q

Which RNA codes for proteins?

A

Only mRNA

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15
Q

True or False:

mRNA is the largest portion of RNA in the cell

A

False, it is the smallest portion of RNA in the cell

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16
Q

How does information transfer occur?

A

Information in DNA is transribed into RNA and then mRNA is translated into proteins

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17
Q

What proteins are involved in each step of information transfer?

A

Enzymes

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18
Q

What shape is DNA? How is this achieved?

A

A double helix, formed by hydrogen bonding

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19
Q

What shape is RNA? How does it achieve this?

A

Can be various shapes, can bond with itself using hydrogen bonding to form these shapes

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20
Q

What shapes are proteins?

A

Varies; depends on function

21
Q

What is generally thought to have evolved first?

A

RNA

22
Q

Define:

Ribozyme

A

A class of catalytic RNA molecules

23
Q

What are two examples of ribozymes?

A

Ribosome: Key organelle that plays a role in the synthesis of proteins
* 2/3 rRNA and 1/3 Protein

Spliceosomes: Involved in removing introns

24
Q

Define:

Transcription

A

DNA to RNA

25
Q

Define:

Translation

A

mRNA to Protein

26
Q

What are the 2 ways to measure gene expression?

A
  1. Transcript Abundance: How much mRNA corresponding to a specific gene is in the cell
  2. Protein Abundance: How much of a protein is in the cell
27
Q

How is transcript abundance measured?

A

Northern Blots (RNA blot hybridization)
Transcription rate and rate of mRNA breakdown

28
Q

How is protein abundance measured?

A

Western Blots
Translation rate and rate of protein breakdown

29
Q

State the similarities and differences between:

DNA and RNA

A

Similarities
* Polymers of nucleotides
* Carriers of information

Differences
* DNA contains a deoxyribose sugar; RNA contains a ribose sugar
* DNA does not have OH at carbon 2
* RNA is less stable and therefore more reactive
* RNA is also attacked by enzymes called ribonucleases

30
Q

Describe:

A Nucleotide

A

5 Carbon sugar + Phosphate + Nitrogenous base

31
Q

Why is it good that RNA degrades more quickly than DNA?

A

Good temporal regulation
We do not need mRNA present all the time if unnecessary as this is a waste of cellular resources

32
Q

True or False:

RNA degrades quickly, so we need to ensure than RNA samples are intact prior to experimentation

A

True

33
Q

Define:

3 types of expression

A
  1. Induced: Up-regulation in gene expression
  2. Constitutive: Expression of the protein or transcript will always be the same
  3. Repressed: Down-regulation in gene expression
34
Q

State:

The groups of “omics”

A

Genomics
Transcriptomics
Proteonomics
Metabolomics

35
Q

What is the purpose of “omics”?

A

Allows us to look at whole genome, transcriptome, proteome etc.

36
Q

In gel images:

What is the RNA and how can we tell size?

A

The glowing rods are RNA, and the further down the strip the smaller it is

37
Q

How is homology proven using molecular info?

A

Look at the genes that make up each homologous structure

38
Q

State the types of:

Expression at level of transcription

A
  1. Basal expression: Normal level
  2. Increased expression: Transcription factors enhance
  3. Decreased expression: Transcription factors inhibit
39
Q

State the types of:

Stability at level of mRNA

A
  1. Basal stability
  2. Increased stability, factors bind to end of mRNA (stabilizing it)
  3. Decreased stability, factors bind to end of mRNA (destabilizing it)
40
Q

What is the difference between regulation at level of promoter and regulation by stability?

A

Regulation at the level of promoter controls transcription
Regulation by stability is at level of mRNA, independent of transcription

41
Q

What are transcription factors?

A

Proteins, binds to DNA promoter to inhibit/promote expression

42
Q

What binds to the promoter of DNA?

A

RNA polymerase

43
Q

What is the half-life of mammalian mRNA? Protein?

A

mRNA: about 9 hrs
Protein: about 46 hrs

44
Q

What is post-translational modification?

A

Modifications made after translation to make the protein functional

45
Q

What is an example of post-translational modification?

A
  • When opsin binds to retinal, it has been modified
  • Functional rhodopsin protein has formed
  • Remember, retinal is not a protein therefore it was not coded from a gene!
46
Q

How do we link a genotype to a phenotype?

A

We mutate the gene and see what happens to the phenotype

47
Q

Compare:

Forward genetics and Reverse genetics

A

Forward genetics: We start with a phenotype, and identify the underlying gene

Reverse genetics: We start with a specific gene which is altered, and identify the phenotype

48
Q

How do we generate a population of mutants?

A
  • Insert a gene randomly into the genomes of organisms
  • Use methods to get the phenotype of interest
49
Q

How does one prove a gene is responsible for a phenotype?

A

After conducting knocking out gene, we rescue the mutant by putting the wild-type copy of the gene back into the mutant
If that gene is responsible for the mutant phenotype, we should get a wild-type phenotype when we rescue it

My Packs: UWO (BIO 1002) (10 decks)

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