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33 Cards in this Set
- Front
- Back
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What causes cell disruption in DNA extraction?
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Physical disruption and detergents.
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What causes the denaturation of proteins in DNA extraction?
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Organic solvents and enzymes.
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What compound is used to precipitate DNA in the extraction process?
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Ethanol.
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What is used to cut DNA in vitro?
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Restriction enzymes.
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What are the DNA joining enzymes?
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Ligases
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What are the DNA synthesis enzymes?
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Polymerases
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Nucleases are used in vitro to...
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Degrade the DNA.
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How do you recognize restriction enzymes?
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They usually create palindromes.
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What is the order of reagents in a restriction enzyme digest?
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Water, 10X salts, DNA, then enzyme.
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What is the melting temperature rule for primer design?
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55-65 degrees C
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What do you have to avoid in primer design?
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Complementarity.
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What is an Ori?
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Origin of replication, unique DNA sequence recongnised by ORC protein complex.
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What does helicase do?
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Unwinds the helix ahead of replication fork
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What does topoisomerase do?
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Untangles the supercoiling
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Flip flopping transcription occurs in?
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Prokaryotes
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What is the Shine-Dalgarno sequence and why is it a core concept?
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AGGAGG used for the 30S ribosomal unit to locate and bind to the nearest AUG codon downstream.
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What does the σ unit do for polymerase in E. Coli?
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Binds the RNA poly to promoter sequence.
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Why are ATT's used in the -10 sequence?
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Less H bonds, therefore easier for σ factor to break open and bind too.
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What is the promoter recognised for in σ 32?
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Heat shock.
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What is the promoter recognised for in σ 28?
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Motility.
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What is the promoter recognised for in σ 54?
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Nitrogen metabolism.
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What is the -35 sequence for σ?
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AAC TGT
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Is Tx on or off in an inducible system?
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Off.
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Is Tx on or off in an repressible system?
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On.
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Does the cAMP have to bind to the CAP before it can bind to the repressor?
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Yes!
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What is required for RNA Pol II action?
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Basal transcription factors.
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What are the two functional domains of TF's?
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DNA binding domain and Activation domain
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What is the funtion of Chromatin?
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To package DNA into smaller volumes
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Is environmental stimuli a main factor in gene control?
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No, it is an exception.
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What edits out introns?
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Spliceosomes.
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What does mRNA stability determine?
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How quickly protein synthesis from its transcription can be shut down.
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What does eIF2:GTP control?
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Initiation of translation. Binds tRNA to ribosomal subunit.
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What two states can eIF2-GDP be in?
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Dephosphorylated (allows tsltn) or Phosphorylated (tsltn will cease)
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