Recombinant DNA Technology Flashcards
What is the significance of the enzyme Taq polymerase? What technique is it used for?
Acts as the DNA polymerase in PCR because it can handle the high temps in the process
define cDNA
the DNA strand made by reverse transcriptase
How are synthetic nucleic acids used?
used to make a DNA or RNA molecule using the precursors and enzymes. Used for determining a genome sequence, synthesizing a gene based on a protein sequence, synthesizing probes to isolate a specific sequence, and synthesizing mRNA that codes for a viral protein.
Define RFLPs
restriction fragment length polymorphism. a collection of DNA fragments that are produced when treating DNA with restriction enzymes
Define GMOs
an organism who’s genetics have been modified to improve it.
Define exons
the protein-encoding parts of DNA
How are gene libraries created? How is this process different for eukaryotic gene libraries?
the genome of an organism is split up gene by gene and put into vectors. Eukaryotic gene libraries are created from cDNA from mRNA to make sure we only get exons
Define Southern blotting
can be used to probe a particular sequence of DNA. DNA is cut using restriction enzymes and then placed in an electric field in gel and fragments separate by length. Fragments are transferred to a nitrocellulose and tested for hybridization with a tagged probe.
Define gene library
a collection of vectors containing single genes
Briefly describe how to sequence DNA. [dideoxy (Sanger) sequencing especially]
copies of the DNA are made w/ four different solutions each containing a small amount of each ddATP (ATGC versions). Those ddATP bind to their corresponding base. This allows us to see where each base is in the sequence.
What is a palindrome?
read the same back as forward
Describe some applications of recombinant DNA technology.
medicine, agriculture, police work, legal stuff
define introns
the non protein-encoding parts of DNA
What are some typical products produced by recombinant DNA procedures?
foods, vaccines, antibiotics, hormones, proteins
Describe four medical products that are genetically engineered.
insulin, tPA, aplha interferon, and hep b vaccine.
Define PCR
a tool used to make copies of DNA, amplify the DNA
What is the role of the vector in recombination?
a DNA molecule (often plasmid or virus) that is used as a vehicle to carry a particular DNA segment into a host cell as part of a cloning or recombinant DNA technique
Define DNA fingerprinting
Originally was mattching patterns of bands that corresponded with RFLPs. Used for crime things and paternity.
Describe 5 ways of introducing DNA into cells.
transformation- naked DNA
electroporation- holes in cell wall
protoplast fusion- cell wall removed by lysozyme and cell membranes merge
gene gun- DNA coated bullets shot into plant cell walls
microinjection- microscopic glass pipets inject DNA into cell
What is reverse transcriptase used for?
coverts mRNA back into cDNA
What is the role of the restriction enzyme in technology?
cuts the DNA into pieces
What is meant by “sticky ends”?
a shape of cut that restriction enzymes make that allows new bases to create hydrogen bonds with the old bases
