8.1 Manipulating & Cloning DNA Flashcards
genetic engineering
intentional production of new genes
and alteration of genomes by the substitution or introduction of
new genetic material
recombinant DNA
fragment of DNA composed of two
sequences originating from at least two different sources
tools for producing recombinant DNA
In order to form recombinant DNA, a molecular biologist
requires tools to cut, join, and replicate DNA
These include:
Restriction enzymes
DNA ligase
Plasmids
restriction enzymes
An enzyme that cuts DNA at specific base pair sequence
recognition site
Each type of restriction enzyme is a able to recognize a specific sequence of nucleotides
restriction fragment
A fragment that is produced when a DNA strand is cut by
these enzymes
function of restriction enzymes
Restriction enzymes are
biological molecules, which are isolated from bacteria
The function of these enzymes in bacteria is to cut foreign viral DNA into pieces
blunt ends
Blunt ends occur when cuts are made straight across the DNA
sticky ends
Sticky ends occurs when cuts are
made in a zigzag across the DNA
DNA ligase
DNA ligase an enzyme that is used for “reassembly” of DNA sticky ends
T4 DNA ligase works well with blunt ends
Joins phosphodiester bonds together
plasmids
Plasmids are small, circular pieces of DNA that are found in
bacteria
A plasmid that has been designed to transfer foreign genetic material into a cell is called a vector
A desired gene must be inserted into a specific location
The same restriction enzyme is used both on the source DNA to isolate target DNA and the plasmid so that they will have the same sticky ends.
restriction maps
Restriction maps or commonly called plasmid maps are a
diagram that illustrates restriction enzyme recognition sites
and the distances, measured in base pairs, between sites.
transformation
Bacteria are able to uptake a plasmids under specific
conditions The introduction of DNA from another source is known as transformation
