How many restriction sites are there for EcoRI?
Note, after a reaction with the EcoRI enzyme, that the DNA of species A is cleaved into three fragments, corresponding to two EcoRI restriction sites, whereas that of species B is cleaved into four fragments, corresponding to three EcoRI restriction sites.
What does EcoRI recognize?
EcoRI recognizes the sequence GAATTC, and cuts both DNA strands between the G and the A nucleotides. Protruding from the cut ends will be single-stranded DNA “tails” having the sequences AATT. Although the nucleotides at the ends of the plasmid and foreign DNA now abut each other, they are not covalently connected.
What are compatible restriction enzymes?
As unlikely as it may seem, restriction enzymes from different organisms can produce interlocking pieces of DNA – so called compatible cohesive ends (CCE). These are pieces of DNA, which fit together and can be ligated, creating a hybrid molecule.
Which enzymes produce compatible sticky ends?
New restriction sites can be generated by ligation of DNA fragments with compatible cohesive or blunt ends. These new restriction sites may be generated by: Cleavage followed by fill-in of 5´ overhangs to generate blunt ends….Compatible Cohesive Ends and Generation of New Restriction Sites.
| Enzyme | Ligated To | Recleaved By |
|---|---|---|
| AgeI * (A/CCGGT) | NgoMIV | BsrFI, HpaII |
How many recognition sites are there in pbr322?
The plasmid has unique restriction sites for more than forty restriction enzymes. Eleven of these forty sites lie within the TetR gene.
How many restriction sites do plasmids have?
A multiple cloning site (MCS), also called a polylinker, is a short segment of DNA which contains many (up to ~20) restriction sites – a standard feature of engineered plasmids. Restriction sites within an MCS are typically unique, occurring only once within a given plasmid.
Does ecor1 produce blunt ends?
EcoRI (pronounced “eco R one”) is a restriction endonuclease enzyme isolated from species E. coli. Other restriction enzymes, depending on their cut sites, can also leave 3′ overhangs or blunt ends with no overhangs.
How does EcoRI specifically act on DNA molecule?
EcoRI enzymes are the class of endonucleases which cut and degrade at the specific position of a DNA by selectively inspecting the length of the DNA sequence. After recognizing the specific sequence, it binds to the site and cuts the DNA strands at sugar phosphate bonds.
Does EcoRI produce sticky ends?
EcoRI creates 4 nucleotide sticky ends with 5′ end overhangs of AATT. Other restriction enzymes, depending on their cut sites, can also leave 3′ overhangs or blunt ends with no overhangs.
Does HindIII create sticky ends?
HindIII restrictions process results in formation of overhanging palindromic sticky ends.