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bt > Gene Design > Flashcards

Gene Design Flashcards

1
Q

dicot or monocot

tobacco

A

Dicot

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2
Q

dicot or monocot

Cotton

A

Dicot

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3
Q

dicot or monocot

Soybean

A

Dicot

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4
Q

dicot or monocot

Legumes

A

Dicot

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5
Q

dicot or monocot

Arabidopsis

A

Dicot

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6
Q

dicot or monocot

Wheat

A

monocot

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7
Q

dicot or monocot

corn

A

monocot

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8
Q

dicot or monocot

Rice

A

monocot

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9
Q

dicot or monocot

Barley

A

dicot or monocot

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10
Q

how does agrobacterium mediated transfer work

A

Ti plasmid to allow transfer of T-DNA section containing transgene to plant, bacteria incubated with leaf discs

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11
Q

Cry1

A

Lepidoptera

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12
Q

Cry3

A

Coleoptera

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13
Q

Cry2

A

Diptera

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14
Q

What species produces Cry toxins

A

Bacillus thuringiensis

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15
Q

When was Bt a registered pesticide

A

1961

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16
Q

Cry toxins have a what structure?

What are the domains?

A

Conserved structure 600aa
3 distinct domains
1-N terminus 7alpha helix membrane insertion pore formation

2- B prism, receptor recognition and binding

3- C terminal antiparallel B sheets receptor recognition and pore formation

17
Q

Digestion of Cry toxins does what

A

Activates then

Digestion to N terminal 60-70kDa truncated form

“Toxin forms lytic pores in cell membrane of insect guy and once digested (active) indigested (inactive)

18
Q

The toxin binds where?

This process is called ?

A

To the high affinity receptors in midgut epithelial

  • forms pores
  • kills e cell

Colliod osmotic lysis

19
Q

What is a protoxin

A

Proteolyticall cleaved/inbound to receptor = not toxic “protoxin”

NOT SURE TBH Might he wrong

20
Q

Lepidoptera

A

Butterflies and moths

21
Q

Diptera

A

Flies and mosquitoes

22
Q

Coleoptera

A

Beetles and weevils

23
Q

Hymenoptera

A

Wasps and bees

24
Q

Herbicide res Maize

Round up

A

Maize = monocot

P - Ubi
FG - aroA (glyphosate red, detoxifies)
T - Nos (nopaline synthase)
MG - nptII kanomycin res

Says for cereals such a maize better to use herbicide selection gene - eg Hydromycin bar gene

25
Q

Aluminium tolerant legumes

A

P- CamV35s
FG- Csb (citrate synthase gene from pseudomonas) - citrate increases too by chelating Al3+
T- NOS nopaline synthase terminator
M- NptII neomycin

Gene construct in cytoplasm not mitochondria so doesn’t interfere with Krebs cycle

26
Q

Butterfly insecticide res in rice

A

Rice = monocot
P -Ubi
FG - Cry1 from bt
T - Nos nopaline synthase term

P-Ubi
FG- CPT1A (protease inhibitor)
T-Nos

P-Ubi
MG- nptII rice :( kanomycin therefore use paromomycin
T-Nos

27
Q

Herbicide too cotton

A

Cotton = Dicot

P-CamV35S
FG - bar (against glufosinate)
T- nos

P-CamV35S
MG- nptII kanomycin
T-nos

28
Q

Tobacco drought/salinity tol

A

P- SARK (senescence associated receptor protein kinase) promoter
TG- IPT (isopentyl transferase, cytokinin biosyn, prevent senescence)
T-Nos nopaline syn term

P-CamV35s
MG- nptII
T-Nos

29
Q

Soybean drought tol

A

P-CamV35s
FG- AtZEP (zeaxanthin epoxidase gene makes zeaxanthin precursor to ABA)
T- Nos

P-CamV35s
FG-nptII kanomycin
T-nos

30
Q

Example of light inducible promoter

A

Promoter from cab gene

31
Q

Use what if don’t what expression all over the plant?

A

Signal transit peptides

Can be used to direct the gene construct to the desired part of the plant
E.g. chloroplast or endosperm

Endosperm useful in rice as a lot of metabolic activity takes place here

32
Q

Drought tol maize

A

P-Ubi
TG- AVP1 (arabidopsis H+-Ppase pump)
T-nos

P-Ubi
MG- bar (herbicide selection w/ PPT)

33
Q

what is a newly developed stacking system

A

Zhu et al (2017) have developed a transgene stacking vector system (TGSII) which allows the successful transformation of plants (tested in rice) with several genes in one construct, each with their own promotors and terminators.

34
Q

How does the TGSII stacking system work

A

These utilise antibiotic selection cassettes (nptII) and Cre/loxp-mediated constructs which allows the marker cassette to be removed under tissue-specific promotors, once the transformants have been selected. This allows a marker free gene construct in planta and it also works with agrobacterium transformation. This could be an interesting alternative for a company to try, as it has had proven success with crop such as Astaxanthin bio-engineered rice (Zhu et al. 2018).

bt (4 decks)

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