Recombinant DNA Flashcards
What is a clone?
An exact genetic copy
What is recombinant DNA?
The joining of a DNA sequence to DNA originating from another source
What is the basis for most recombinant DNA techniques?
Complementary base pairing
What tools are needed for cloning?
Source of DNA
Vector
Several bacterial enzymes and processes
Selection methods
What vectors can be used to create recombinant DNA?
Plasmid
Phage
Cosmid
YAC
Forensic analysis uses what type of information?
Simple repeats (microsatellites)
What is cDNA synthesis?
Using a retroviral enzyme to make DNA copies of mRNA
What retroviral enzyme is used in cDNA synthesis?
Reverse transcriptase
What is the process of cDNA synthesis?
Beginning with an mRNA, a polyT complement the the polyA tail is formed to be a primer for reverse transcriptase
The DNA copy is then made from RNA
What is the easiest vector to use for cloning?
Plasmids
What limits the usefulness of plasmids for cloning?
Limited in how much information can be inserted
Which of the 4 vector types is rarely used in cloning?
Cosmids
What is the important plasmid to remember?
pBR322
What are the resistance sites found on the pBR322 plasmid?
Ampicillin resistance (ampr) Tetracycline resistance (tetr)
What is the cutting site for the ampr portion of pBR322?
Pstl
What are the cutting sites for the tatr portion of pBR322?
BamHI
SalI
What are the two cycles that a bacteriophage can undergo?
Lytic cycle
Lysogenic cycle
The lysogenic cycle results in what?
Production of dormant bacteriophage
What are type II restriction endonucleases?
Bacterial defense mechanism
What do type II restriction endonucleases recognize?
4-8 base pair palindromes
Type II restriction endonucleases may produce what?
Sticky ends
What is used to fragment plasmid and human DNA so that they have sticky ends?
Restriction enzyme
If a phage in a soft agar lyses the cells around it, what does it cause?
Plaques
Probes are used in what reactions?
Hybridization reactions
Gene therapy has been used to treat what ocular condition?
Choroideremia - to prevent blindness
What does a pluripotent cell come from?
Blastocyst
What can a pluripotent cell form?
Cell types from all 3 germ layers
Where does a multipotent cell come from?
An adult
What can a multipotent cell form?
Closely related cell types
What is ARMS PCR used for?
To detect point mutations using allele-specific primers
