Correct answers
5 procedures in cloning Correct Answer--obtaining DNA to be cloned
-selecting a vector
-joining the vector and DNA, using ligase to seal
-transform a host organism
-select host cells that have cloning vector
Correct Answer--made by bacteria
-cleave DNA at specific recognition sites
plasmid Correct Answer--aka a vector
-a circular DNA molecule that reproduces separately from the
chromosome, contains nonessential gens picked up from the
environment
transformation Correct Answer-the process by which plasmids are
introduced into bacterial cells
selectable marker gene Correct Answer--used to tell which bacterial
cells have picked up the plasmid
-eg antibacterial resistance
, Origin of replication Correct Answer-sequence in the plasmid where
replication is initiated
-required to propagate the plasmid
polylinker Correct Answer-place near promoter on a plasmid where the
GOI is inserted
transcription-termination sequence Correct Answer-a sequence that can
improve the quality and amount of mRNA produced
Why does translation need to be regulated during gene cloning? Correct
Answer-If the cell makes too much of the protein, the increased
concentration of protein can kill the cells
oligonucleotide directed mutagenesis Correct Answer--use a plasmid
with a targeted start site fo the mutation
-denature and add primers with the mutation in them
-use DNA pol to replicate plasmid and incorporate primer inside
-digest non mutated parent DNA templates
-transform dsDNA (w/mutant)
How are primers synthesized Correct Answer--first nucleoside is bound
to silica support
-next nucleotide is added
-repeat until all nucleotides are added