Term
| The real purpose of restriction enzymes is _____________________ |
|
Definition
| to protect bacteria from invading DNA |
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Term
| Restriction enzymes have been very important to _________ |
|
Definition
| manipulate, move, and propagate pieces of DNA in E.coli using plasmid cloning vectors. |
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Term
| Why don't bacteria digest their own DNA with restriction enzymes? |
|
Definition
Restriction enzymes typically come from two genes or
activities, a restriction activity and a methylase activity.
Methylation of DNA bases at the recognition sequence
prevent the RE from cutting. Since entering
bacteriophage DNA is not methylated, it is cut. |
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Term
| Type II and Type II enzymes cleave DNA chains at ________. |
|
Definition
| specific recognition sequences |
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Term
| How many bases do enzymes usually recognize? |
|
Definition
|
|
Term
| How much ATP is required for digestion? |
|
Definition
|
|
Term
|
Definition
E=E.
co = coli
RI = 1st restriction enzyme found in R strain of E. coli |
|
|
Term
| EcoRI sequence recognized |
|
Definition
|
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Term
| If you start with a complex DNA, like human genomic DNA isolated from cells, the digestion products will _____ the gel. |
|
Definition
| appear as a smear from top to bottom of |
|
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Term
|
Definition
| pairs of restriction enzymes specific to the same recognition sequence |
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Term
|
Definition
| developed the dideoxy method and found the aa sequence for insulin (2 nobel prizes) |
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Term
| dideoxynucleotide analogs |
|
Definition
nucleotide analogs used as chain terminators to determine sequence of DNA
They possess no 3'-OH. Instead they have a 3'-H that cannot be used for elongation. |
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|
Term
| dideoxynucleotide electrophoresis sequence is read ____________________ |
|
Definition
| from the bottom of the gel to the top -> this is the complement of the template. |
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Term
|
Definition
Template DNA molecules are immobilized on microscopic beads. Hundreds of thousands to millions of beads are analyzed at once so that sequencing of many different DNAs are performed and
read at the same time. |
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Term
|
Definition
This is released whena nucleotide is correctly added to the end of one of
the DNA templates. This is used in 454 Sequencing. |
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Term
| What enzymes work with pyrophosphate to produce light? |
|
Definition
| ATP sulfurylase and luciferase |
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Term
| Reaction catalyzed by ATP sulfurylase |
|
Definition
| PPi + APS → ATP + SO42- (used in conversion of PPi to light) |
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|
Term
| Reaction catalyzed luciferase |
|
Definition
| ATP + luciferin + O2 → AMP + CO2 + oxyluciferin + light |
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