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Bio 361 - Molecular Genome and Genomics > Midterm 2 - Notes 6 (Part 7) > Flashcards

Midterm 2 - Notes 6 (Part 7) Flashcards

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Biology 101 flashcards
1
Q

What are plant transformation goals?

A

All cells of a plant should contain the new construct

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2
Q

What is a problem in plant tranformation?

A

Only identical cells are being transformed to received the T-DNA

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3
Q

What is plant transformation?

A

Delivery of DNA to a single plant cell

  • integration of DNA into the genome
  • conversion of transformed cell into a whole plant
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4
Q

What are all plant cells?

A

Totipotent

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5
Q

Totipotent

A

Can regenerate complete plants when provided with nutrients and growth hormones

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6
Q

What do reporter gene constructs follow?

A

The organ, tissue and cell type of specific expression of YGI

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7
Q

What do reporter gene constructs isolate?

A

The promoter of YGI

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8
Q

What are 2 examples of a reporter gene?

A
  1. GUS

2. LUC

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9
Q

What does GUS encode?

A

A beta-glucuronidase that cleaves a colourless substrate (X-Gluc) to produce a blue precipitate

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10
Q

What does LUC encode?

A

A firefly enzyme (luciferase) that oxidizes luciferin and thereby emits light (luminescence)

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11
Q

What does YGI reporter gene fusion constructs follow?

A

The sub-cellular localization of YGI

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12
Q

What are the 5 steps involving YGI-reporter gene fusion constructs?

A
  1. Follow the sub-cellular localization of YGI
  2. Isolate the open reading frame of YGI
  3. Fuse it to a reporter gene
    - GFP
  4. Fuse construct to a constitutive promoter
  5. Visualize fluorescence in vivo using confocal laser microscopy
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13
Q

What results would you get?

A

Would get both expression, where it is normally expressed in sub cellular expression

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14
Q

What can over expressed constructs identify?

A

GOF and LOF

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15
Q

Can cDAN of YGI come from any organism?

A

Yes

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16
Q

What kind of marker is knock down constructs?

A

Always a selective marker between the LR and the RB

- located towards the IB

17
Q

Where does T-DNA degration start?

A

At the RB and the LB is integrated last
- if you get selection marker present you can be sure the construct of integration is first and the selection marker follows that

18
Q

What is the knock down constructs used for?

A

To abolish/reduce expression of YGI

19
Q

What are the 5 steps in knock down constructs (anti-sense)?

A
  1. Isolate outer reading frame of YGI
  2. Clone it in inverse orientation in from of a promoter (anti-sense)
  3. Or clone parts of it twice in reverse orientation, separated by an intron (RNAi)
  4. Transform the plant of origin to YFG
  5. When transcribed, the transgene produces an anti-sense mRNA (complementary to the endogenous sense-mRNA from YGI)
20
Q

What are 3 promoter choices for knock down constructs?

A
  1. Constitutive
    - cauliflower
  2. Inducible promoter
  3. YGI promoter
21
Q

What are 3 promoter choices for over expression constructs?

A
  1. Constitutive
    - cauliflower
  2. Inducible promoter
  3. Tissue/cell style promoters
22
Q

What is an example of transgenic plants used in food production?

A

Herbicide resistant plants

23
Q

What are herbicides normally?

A

Non selective

- can kill both weed and crop plant

24
Q

What do herbicides target?

A

Biological processes that is not important in other organisms groups

25
Q

What are herbicides specific to?

A

Killing plants and not animals

26
Q

When are herbicides usually applied?

A

Before crop plants germinate

27
Q

What are 4 possibilities to engineer herbicide resistant plants?

A
  1. Inhibition of uptake
  2. Add protein that inactivates herbicide
  3. Over production target protein
    - produce way more of the target protein so even though its being killed off there is still enough to function
  4. Replace target protein with resistant variety **
28
Q

What is an example of transgenic plants use in herbicide resistance?

A

Glyphosate resistant plants - roundup ready

29
Q

What pathway is present in plants, bacteria and some fungi, but not animals?

A

Shikimate pathway

30
Q

Why is shikimate pathway specific to plants?

A

Because the pathway does not exist in animals

- this is the reason the aromatic amino acids are essential when taking up our food

31
Q

What does glyphosphate resistant plants lead to?

A

Biosynthesis of essential aromatic amino acids

32
Q

What does glyphosate bind to?

A

Binds to the active site of ESPS synthase and inactivates it

33
Q

What is the mutated ESPS synthase version from E.Coli insensitive to?

A

Glyphosate

  • but still active
  • does not inhibit the version of the enzyme
  • allows them to easily create transgenic plants

Bio 361 - Molecular Genome and Genomics (35 decks)

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