recombinant DNA Flashcards
recombinant protein is one that has been obtained via
transcribing and translating a novel combination of DNA from different organims
an example of recombinant protein is the gene for human insulin being placed in a bacterial plasmid which allows the plasmid to?
produce insulin that can be used to treat diabetes
restriction endonucleases are bacterial enzymes that recognize specific?
sequences of DNA and cut dsDNA into two pieces
nuclease is an enzyme that
cuts nucleic acids thus endonuclease cuts in the middle of a DNA chain
sticky ends of DNA are
complementary free ends of a DNA molecule that are able to base pair with other DNA molecules with similar sequence
palindromes are sequences with
two-fold rotational symmetry
when a fragment of dsDNA is created by cutting with a restriction endonuclease, it can be inserted to?
DNA from another source that was also digested by the same restriction endonuclease
plasmids are?
small circular dsDNA molecules found in bacteria and are capable of autonomous replication
autonomous replication is
replication that is independent of chromosome replication
plasmids can be manipulated by recombinant techniques to?
propagate and express foreign genes in bacteria
plasmids have multiple cloning sites, which has?
restriction sites for dozens of restriction enzymes
plasmids also have drug resistance gene that helps them?
select and isolate bacteria possessing the plasmid from other bacteria
bacteria containing plasmid with ampicillin-resistance gene are able to grow in the presence of?
of the antibiotic ampicillin
bacterial transformation is when plasmids are?
reintroduced back into bacterial cells
plasmids are taken back into bacterial cells by?
cooling cells in CaCl2 and then heat shocking them
&
electroporation which is when an electric field is applied and pokes holes in membrane
once plasmid is reintroduce to bacterial cell, it will
be exposed to host’s replication machinery which replicates the plasmid
complementary DNA is produced from?
fully spliced eukaryotic mRNA by enzymes that read RNA template and builds cDNA
cDNA can be ligated into plasmids thus
bacteria will produce the protein encoded by the cDNA
cDNA libraries are compared across tissue type of organism in research to?
study tissue-specific gene expression
bacterial artificial chromosomes (BACs) can insert 100 to 350 kilobase pairs and contain sequences that allow for?
replication and regulation of copy number, partition genes that promote their even distribution after bacterial cell division, and a selectable marker for antibiotic resistance
yeast artificial chromosomes (YACs) can insert 100 to 3,5000 kilobase pairs and contain?
a centromere, telomeres, and sequences that function as replication origins
BACs are used to study?
inherited diseases (Alzheimer's and Down Syndrome) that involve complex genes with several regulatory sequences and promoters upstream of the coding sequence BACs can clone entire genome of some viruses such as herpes, poxviruses, and coronaviruses
