Term
| 5 Steps to Recombining Molecules |
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Definition
1. Must be ale to cut and rejoin DNA in precise locations. Use sequence specific endonucleases
2. Select DNA molecule to serve as a carrier
3. Prepare and insert foreign DNA, vector with inserted DNA known as recombinant DNA
4. Introduce vector/insert into host organism
5. Screen for host cells replicating the hybrid DNA |
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Term
| Type II Restriction Endonuclease |
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Definition
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Term
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Definition
| Covalent joining of free DNA ends |
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Term
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Definition
| Copy DNA single strand to dsDNA |
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Term
| Type II Restriction Endonuclease |
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Definition
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Term
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Definition
| Phosphorylates 5'-OH group for labeling or ligation |
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Term
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Definition
| Homopolymerization of tails to 3'-OH ends |
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Term
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Definition
| Deletes bases from 3' ends of DNA strand |
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Term
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Definition
| Removes 5'-3' single strand to produce ss 3' overhang |
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Term
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Definition
| Removes terminal phosphates from ends |
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Term
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Definition
Circular dsDNAs with 3 essential components:
A replicon, a selectable gene, an expression locus |
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Term
| Amplification of Clone in Host |
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Definition
| Cloning vector inserted into host organism, organism allowed to grow/dna duplicates thus amplifies the clone, must identify cells with cloning vector. |
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Term
| Transformation efficiency |
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Definition
| # of tetR colonies/ # of total viable cells plated |
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Term
| Building DNA Library Shotgun Approach Equation |
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Definition
P= 1-(1-f)^N
N= (ln (1-p))/(ln(1-f))
Use to calculate the number of clones required to have prob P of isolating a particular gene. |
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Term
| Site-Directed Mutagenesis |
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Definition
| DNA is altered by modifying nucleotide sequence in order to change the amino acid sequence of the encoded protein. Synthetic oligonucleotides required to specifically alter the gene. They hybridize to the DNA around the area with the desired new base sequence. |
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Term
| Transgenic Organism (animals and plants) |
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Definition
Express a foreign gene or transgene
Allows the engineering of an animal as an experimental model for a human disease.
Allows the expressions of proteins needed for better nutrition |
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Term
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Definition
Introduction of a gene order to produce a therapeutic effect
Various gene therapy trials ongoing
Only documented success thus far is for severe combined immunodeficiency disease. |
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