Restriction Enzymes Molecular Scissors Restriction enzymes are molecular

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Restriction Enzymes

Restriction Enzymes

Molecular Scissors Restriction enzymes are molecular scissors

Molecular Scissors Restriction enzymes are molecular scissors

• Restriction Enzymes scan the DNA code • Find a very specific set

• Restriction Enzymes scan the DNA code • Find a very specific set of nucleotides • Make a specific cut

Picking a palindrome Words that read the same forwards as backwards Hannah hanna. H

Picking a palindrome Words that read the same forwards as backwards Hannah hanna. H Level leve. L Madam mada. M

Palindromes in DNA sequences Genetic palindromes are similar to verbal palindromes. A palindromic sequence

Palindromes in DNA sequences Genetic palindromes are similar to verbal palindromes. A palindromic sequence in DNA is one in which the 5’ to 3’ base pair sequence is identical on both strands (the 5’ and 3’ ends refers to the chemical structure of the DNA).

Each of the double strands of the DNA molecule is complimentary to the other;

Each of the double strands of the DNA molecule is complimentary to the other; thus adenine pairs with thymine, and guanine with cytosine.

Restriction enzymes recognize and make a cut within specific palindromic sequences, known as restriction

Restriction enzymes recognize and make a cut within specific palindromic sequences, known as restriction sites, in the genetic code. This is usually a 4 - or 6 base pair sequence. Example?

Hae. III is a restriction enzyme that searches the DNA molecule until it finds

Hae. III is a restriction enzyme that searches the DNA molecule until it finds this sequence of four nitrogen bases. 5’ TGACGGGTTCGAGGCCAG 3’ 3’ ACTGCCCAAGGTCCGGTC 5’

Once the recognition site was found Hae. III could go to work cutting (cleaving)

Once the recognition site was found Hae. III could go to work cutting (cleaving) the DNA 5’ TGACGGGTTCGAGGCCAG 3’ 3’ ACTGCCCAAGGTCCGGTC 5’

These cuts produce what scientists call “blunt ends” 5’ TGACGGGTTCGAGG 3’ ACTGCCCAAGGTCC CCAG 3’

These cuts produce what scientists call “blunt ends” 5’ TGACGGGTTCGAGG 3’ ACTGCCCAAGGTCC CCAG 3’ GGTC 5’

The names for restriction enzymes come from: • the type of bacteria in which

The names for restriction enzymes come from: • the type of bacteria in which the enzyme is found • the order in which the restriction enzyme was identified and isolated. Eco. RI for example R strain of E. coli bacteria I as it is was the first E. coli restriction enzyme to be discovered.

“blunt ends” and “sticky ends” Remember how Hae. III produced a “blunt end”? Eco.

“blunt ends” and “sticky ends” Remember how Hae. III produced a “blunt end”? Eco. RI, for instance, makes a staggered cut and produces a “sticky end” 5’ GAATTC 3’ 3’ CTTAAG 5’ 5’ G AATTC 3’ 3’ CTTAAG 5’

blunt end sticky end

blunt end sticky end

Some more examples of restriction sites of restriction enzymes with their cut sites: Hind.

Some more examples of restriction sites of restriction enzymes with their cut sites: Hind. III: 5’ AAGCTT 3’ 3’ TTCGAA 5’ Bam. HI: 5’ GGATCC 3’ 3’ CCTAGG 5’ Alu. I: 5’ AGCT 3’ 3’ TCGA 5’

“sticky ends” are useful DNA fragments with complimentary sticky ends can be combined to

“sticky ends” are useful DNA fragments with complimentary sticky ends can be combined to create new molecules which allows the creation and manipulation of DNA sequences from different sources. Think about how this could be used and abused in the medical field

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