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23 Cards in this Set

  • Front
  • Back
what is the main goal of stable transfection?
to permanently introduce a gene to the genome of a cell
what sort of selectable marker is used in transfection to isolate cells that take up the gene of interest?
antibacterial resistance
what is the primary disadvantage of stable transfection?
it is very time-consuming to create a stable cell line
what is the main advantage of transient transfection?
the procedure is quick
does the plasmid in transient transfection replicate with the host genome? Why or why not?
no; it does not have a eukaryotic origin of replication
what is the common drawback of both stable and transient transfection?
only a small percentage of cells will take up the DNA -- the process is inefficient
in addition to a bacterial origin of replication and gene for antibacterial resistance, what must be found on gene to be introduced via transfection?
eukaryotic enhancer
in what form is DNA introduced in stable transfection? Through which method is the DNA introduced into genome?
linear DNA; non-homologous end joining.
what aspect of transfected DNA is required in stable transfection but not in transient transfection?
resistance gene for growing eukaryotic cells
what proteins are encoded by the gag gene in retroviruses?
viral capsid proteins
which enzymes are encoded by the pol gene in retroviruses?
reverse transcriptase, integrase, and protease
which proteins are encoded by the env gene in retroviruses?
transmembrane and receptor-binding proteins
what is the role of the cis-acting sequences PB and psi in the retroviral genome?
PB is the binding sites for primers for reverse transcriptase; psi interacts with capsid proteins to allow efficient incorporation of the newly synthesized retroviral genome into viral particle
what is the function of sequences at the end of the long terminal repeats?
to help with insertion of the ds DNA genome into host cell chromosomes by integrase
in a retroviral vector, which genes of the retrovirus have been replaced by the gene of interest?
env, pol. Gag
what is the name for the virus once its genome has integrate into that of the host?
what is the nature of the retroviral genome?
ss RNA
how are viral proteins obtained in a retroviral vector?
they are added in trans
what is the name for the line of cells which has been programmed to express the gag, env, and pol proteins or retroviruses?
packaging cell line
what is the unique advantage of having the pol, gag, and env genes on a genome apart from the retroviral genome?
in subsequent progeny, these proteins will not be expressed and transfection will not continue indefinitely
what is one serious problem with retroviral infections?
they insert their genome anywhere in the cell. If the genome interrupts a host gene which encodes a protein, this can be a major problem
what is a limitation of mouse retroviruses?
they can only infect cells that are dividing bc genome/capsid is otherwise too large to enter nucleus.
what is the upper limit of the size of genes of interest allowed to be transfected via retroviruses?