Table of Contents
What is the recognition sequence for EcoRI?
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.
What is the recognition sequence for 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 the recognition sequence of Hind 2?
Hind II recognizes the sequence GTPy/PuAC and generates fragments with blunt ends (1). Compatible ends Hind II generates fragments with blunt ends and is compatible to any other blunt end. Isoschizomers Hind II is an isoschizomer to Hinc II.
What is recognition endonuclease sequence?
Most restriction endonucleases will reproducibly cleave DNA at a precise point within a recognition sequence. Generally, different enzymes will recognize different sequences that are four to six nucleotides long.
What does EcoRI signify?
EcoRI signifies the name of restriction endonuclease derived from bacterium Escherichia coli. (i) First capital letter of the name comes from the genus of the organism Escherichia is ‘E’. (ii) Second two small letters comes from the species of cells from which they are isolated i.e. ‘co’.
What do you mean by recognition sequence?
A recognition sequence is a DNA sequence to which a structural motif of a DNA-binding domain exhibits binding specificity. A recognition sequence is different from a recognition site. A given recognition sequence can occur one or more times, or not at all, on a specific DNA fragment.
How many fragments are produced by BamHI?
BamHI cleaves the same molecule into fragments of sizes 6.0, 10.1, and 12.9, and the label is found associated with the 6.0 and 10.1 fragments. When EcoRI and BamHI are used together, fragments of sizes 1.0, 2.0, 2.9, 3.5, 6.0, 6.2, and 7.4 kb are produced.
What is correct about Hind II?
Hind II was the first discovered restriction endonuclease enzyme. It has been isolated from Haemophilus influenzae Rd. It cuts DNA molecules at a particular point by recognising a specific sequence of six base pairs. This specific sequence is known as the recognition sequence for Hin d II.
What is Type 2 restriction enzyme?
Type II restriction endonucleases are components of restriction modification systems that protect bacteria and archaea against invading foreign DNA. Most are homodimeric or tetrameric enzymes that cleave DNA at defined sites of 4-8 bp in length and require Mg2+ ions for catalysis.
Is BamHI sticky or blunt?
Recognition Sequences
| Enzyme | Organism | Blunt or Sticky End |
|---|---|---|
| EcoRI | Escherichia Coli | Sticky |
| BamHI | Bacillus amyloliquefaciens | Sticky |
| BglII | Bacillus globigii | Sticky |
| PvuI | Proteus vulgaris | Sticky |
What is EcoRI How does it function?
EcoRI (pronounced “eco R one”) is a restriction endonuclease enzyme isolated from species E. coli. It is a restriction enzyme that cleaves DNA double helices into fragments at specific sites, and is also a part of the restriction modification system. EcoRI creates 4 nucleotide sticky ends with 5′ end overhangs of AATT.
Is the target sequence 5’CTGCAG the same as PstI?
Both enzymes recognize the target sequence 5’CTGCAG. The enzyme systems have similar methyltransferases (41% amino acid identity), restriction endonucleases (46% amino acid identity), and genetic makeup (58% nucleotide identity). These observations suggest a shared evolutionary history.
What are the two components of the PstI system?
The PstI restriction/modification (R/M) system has two components: a restriction enzyme that cleaves foreign DNA, and a methyltransferase which protect native DNA strands via histone methylation. The combination of both provide a defense mechanism against invading viruses.
How many base pairs are needed in the PstI system?
In the PstI system, the genes are encoded on opposite strands and hence must be transcribed divergently from separate promoters. The transcription initiation sites are separated by only 70 base pairs.
How is a recognition sequence different from a recognition site?
The restriction endonuclease PstI recognizes, binds, and cleaves the sequence 5′-CTGCAG-3′. A recognition sequence is different from a recognition site. A given recognition sequence can occur one or more times, or not at all, on a specific DNA fragment.