Ch 11 - Genetic Engineering Flashcards
recognize specific DNA sequences and cut DNA at those sites
restriction enzymes
Protect prokaryotes from hostile foreign DNA (e.g., viral genomes)
restriction enzymes
protect cell’s DNA for restriction enzymes
modification enzymes
method used to separate DNA molecules based on size
gel electrophoresis
base pairing of single strands of DNA or RNA from two different sources to give a hybrid double helix
nucleic acid hybridization
a hybridization procedure where DNA is in the gel and probe is RNA or DNA
southern blot
a hybridization procedure where RNA is in the gel and probe is RNA or DNA
northern blot
isolation and incorporation of a piece of DNA into a vector so it can be replicated and manipulated
molecular cloning
What are the three main steps of gene cloning?
1) Isolation of source DNA
2) Insertion of DNA into vector
3) Introduction of cloned DNA to host
enzyme that joins two DNA molecules
DNA Ligase
performed in vitro and introduces mutations at a precise location
site-directed mutagenesis
cassettes are inserted into the middle of the gene
gene disruption
Encode proteins that are easy to detect and assay
reporter genes
Promoters or coding sequences of genes of interest can be swapped with those of reporter genes to elucidate gene regulation under various conditions
gene fusions
Vector transfer is carried out by ______ or ________
chemical transformation or electroporation
in blue/white screening, ______ colonies do not have vector with foreign DNA inserted
Blue colonies
in blue/white screening _______ have foreign DNA inserted
white colonies
lacZ gene is inactivated by insertion of foreign DNA
insertional inactivation
allow experimenter to control the expression of cloned genes
expression vectors
Effective _______ _________ are used to prevent expression of other genes on the plasmid
transcription terminators
