What is the recognition sequence for the restriction enzyme HindIII?
Under the standard reaction conditions, the restriction endonuclease HindIII cleaves double-stranded DNA, within the recognition sequence–A/AGCTT–at the position indicated by the arrow. In the presence of dimethyl sulfoxide the substrate specificity of this enzyme is reduced and cleavages occur at additional sites.
What is restriction enzyme sequence?
A restriction enzyme is a protein that recognizes a specific, short nucleotide sequence and cuts the DNA only at that specific site, which is known as restriction site or target sequence. In live bacteria, restriction enzymes function to defend the cell against invading viral bacteriophages.
How many fragments did restriction enzyme make in HindIII?
After digestion with EcoRI, you obtain four fragments: 1, 2, 3, and 4. After digestion of each of these fragments with HindII, you find that fragment 3 yields two subfragments (31 and 32) and that fragment 2 yields three (21, 22, and 23).
Is HindIII a restriction enzyme?
Endonuclease HindIII is a type II restriction enzyme which recognizes and cleaves the palindromic sequence AAGCTT in the presence of Mg2+.
What is BAM Hind II?
HindIII (pronounced “Hin D Three”) is a type II site-specific deoxyribonuclease restriction enzyme isolated from Haemophilus influenzae that cleaves the DNA palindromic sequence AAGCTT in the presence of the cofactor Mg2+ via hydrolysis.
What is BamHI restriction enzyme?
BamHI (from Bacillus amyloliquefaciens) is a type II restriction endonuclease, having the capacity for recognizing short sequences (6 bp) of DNA and specifically cleaving them at a target site. DNA is bound in a large cleft that is formed between dimers; the enzyme binds in a “crossover” manner.
Is EcoRV a restriction enzyme?
EcoRV (pronounced “eco R five”) is a type II restriction endonuclease isolated from certain strains of Escherichia coli. It has the alternative name Eco32I. In molecular biology, it is a commonly used restriction enzyme.
What is the biological function of a restriction enzyme?
Restriction enzymes are found in many different strains of bacteria, where their biological role is to participate in cell defense. These enzymes “restrict” foreign DNA (e.g. viral) that enter the cell, by destroying it.
Why is my restriction enzyme not cutting DNA?
Restriction enzymes are responsible for cut the DNA at spacefic site. But restriction enzyme can’t cut their own genome or DNA; because bacterial genome has a gene which is known as DAM gene by which a spacefic type of enzyme is produced which is known as methylases which is responsible for…
How many classes of restriction enzymes are there?
Based on the types of sequences identified, the nature of cuts made in the DNA, and the enzyme structure, there are three classes: Type I restriction enzymes, Type II restriction enzymes, and Type III restriction enzymes.
What is an example of restriction enzyme?
SmaI is an example of a restriction enzyme that cuts straight through the DNA strands, creating DNA fragments with a flat or blunt end. Other restriction enzymes, like EcoRI , cut through the DNA strands at nucleotides that are not exactly opposite each other.