Term
| What are some possible control points in gene expression in transgenic plants? |
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Definition
Inside nucleus:
Transcription
RNA processing
Outside nucleus:
RNA transport
Translation
RNA stability
Protein activity
Protein stability |
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Term
| You are introducing a new gene in a plant. You might want to control: |
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Definition
WHERE you want the gene active
WHEN you want the gene active
HOW active you want the gene
HOW long you want it active for |
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Term
| What are cis (upstream of TIC) and trans proteins (downstream) that regulate eukaryotic transcription on the genome? |
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Definition
Trans: activator and repressor proteins
Cis: enhancers and silencers |
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Term
| What are some different kinds of promoters? |
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Definition
Strong
Weak
Constitutive
Inducible
Developmentally regulated
Tissues specific
Time regulated |
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Term
| How would you study a promoter in the lab? |
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Definition
Clone promoter region
Fuse promoter to a reporter gene
Transform plants and study expression in transgenic plants |
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Term
| What are some examples of reporter genes? |
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Definition
GUS (produces blue colour)
GFP (fluorescent)
Luciferase (produces light from firefly) |
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Term
| Why is it difficult to express a foreign gene in plants? What must be done to efficiently express a foreign gene |
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Definition
| Prokaryotes and eukaryotes have different genetic structures, even within eukaryotes (introns and codon usage). To efficiently express a foreign gene it is often necessary to perform a number of modifications in the original gene sequence |
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Term
| What are three methods for knocking out a gene? How affective is each of these methods? |
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Definition
Antisense inactivation: Clone plant gene, flip it around the other way, reintroduce into plant (20 percent of transgenic plants will have endogenous gene silenced)
Sense co-suppression: Clone plant gene, add strong promoter, reintroduce into plant. (Most produce large amounts of product BUT 10 percent will have gene silenced)
RNA interference: Clone plant gene, prepare construct, reintroduce into plant (90 percent of transgenic plants will be silenced) |
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