Term
|
Definition
-form of DNA that does not exist naturally
-created by combining DNA sequences that would not normally occur together
-introduced through the addition of relevant DNA into an existing organismal DNA (such as plasmids of bacteria) to code for or alter different traits for a specific purpose
-does not occur through processes in the cell but is engineered |
|
|
Term
|
Definition
-vector is isolated
-DNA is cut into fragments
-gene inserted into vector
-vector is taken up by cell
-cell with gene of interest are cloned
-copies of gene are harvested/cells make a protein product
-desired proteins are harvested |
|
|
Term
|
Definition
-site directed mutagenesis
-restriction endonucleases
-vectors
-PCR
-cDNA synthsis |
|
|
Term
| site directed mutagenesis |
|
Definition
|
|
Term
| restriction endonucleases |
|
Definition
-enzymes produced by bacteria
-protect from viral infection by destroying viral DNA
-bacteria protect their DNA by methylating some cytosines
-now produced in the lab
-recognize only one particular sequence
-widely used in gene cloning |
|
|
Term
|
Definition
-self replicationg
-suitable size for manipulation (smaller the better)
-preservation (plasmids are circular, viral DNA inserts quickly)
-presence of marker gene |
|
|
Term
|
Definition
-polymerase chain rxn
-allows small samples of DNA to be amplified
-exponential process
-used of DNA polymerase from thermophilic bacteria (Thermus aquaticus)
-amplified DNA can be examined with gel electrophoresis |
|
|
Term
|
Definition
-complementary DNA
-eukaryotes contain exons and introns
-for cloning bacteria, introns must be removed since prokaryotes cant remove them
-using mRNA from gene of interest, reverse transcriptase can synthesize a cDNA stand used for PCR
-a second strand is synthesized using DNA polymerase |
|
|
Term
| how to get foreign DNA into cells |
|
Definition
-transformation
-electroporation
-protoplast fusion
-gene gun
-microinjection |
|
|
Term
|
Definition
| -cells take up DNA from environment naturally or induced (calcium chloride weakens cell membrane, DNA mixed with competent cell-heat shock) |
|
|
Term
|
Definition
| electric current used to create pores in cell membrane |
|
|
Term
|
Definition
-protoplasts are cell without cell wall (removed by enzymatic digestion)
-more direct access to plasma membrane
-widely used in plant and algal genetics |
|
|
Term
|
Definition
-used to introduce DNA into plant cells
-microscopic gold or tungsten particles coated with DNA and shot directly through the cellulose wall |
|
|
Term
|
Definition
| injection of DNA directly into cell using micropipette |
|
|
Term
| how to identify cell that has acquired foreign DNA |
|
Definition
-blue white screening
-colony hybridization |
|
|
