What does EcoRI produce?

EcoRI creates 4 nucleotide sticky ends with 5′ end overhangs of AATT. The nucleic acid recognition sequence where the enzyme cuts is G↓AATTC, which has a palindromic, complementary sequence of CTTAA↓G. 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?

EcoRI is a restriction enzyme that cleaves DNA double helices into fragments at specific sites. The / in the sequence indicates which phosphodiester bond the enzyme will break in the DNA molecule. Other restriction enzymes, depending on their cut sites, can also leave 3′ overhangs or blunt ends with no overhangs.

What DNA sequence does EcoRI bind?

sequence GAATTC
For example, an endonuclease isolated from Escherichia coli, called EcoRI, recognizes the sequence GAATTC, and wherever this occurs in double-stranded DNA, it will cleave between the G and A (see Figure 2-3).

Where does the EcoRI enzyme cut?

EcoRI cuts double stranded DNA at the sequence GAATTC, but note that this enzyme, like many others, does not cut in exactly the middle of the restriction sequence (Figure 8.4. 8). The ends of a molecule cut by EcoRI have an overhanging region of single stranded DNA, and so are sometimes called sticky-ends.

Is EcoRI a cloning vector?

The unique PsH and PvuI restriction sites present in these vectors are located in the ampicillin resistance (Ap r) gene, while the EcoRI site in pBR325 is located in the chloramphenicol resistance (Cm r) gene. This cloning vector, design~.

What is EcoRI How does it function?

EcoRI is a restriction enzyme that cleaves DNA double helices into fragments at specific sites. It is also a part of the restriction modification system. In molecular biology it is used as a restriction enzyme. EcoRI creates 4 nucleotide sticky ends with 5′ end overhangs of AATT.

What sequence does BamHI cut?

BamHI binds at the recognition sequence 5′-GGATCC-3′ , and cleaves these sequences just after the 5′-guanine on each strand. This cleavage results in “sticky ends” which are 4 b.p. long.

Is pBR322 a DNA?

pBR322 DNA is a commonly used plasmid cloning vector in E. coli (1). The molecule is a double-stranded circle 4,361* base pairs in length (2). pBR322 contains the genes for resistance to ampicillin and tetracycline, and can be amplified with chloramphenicol.

How does the restriction enzyme know where to cut the plasmid?

The enzyme “scans” a DNA molecule, looking for a particular sequence, usually of four to six nucleotides. Once it finds this recognition sequence, it stops and cuts the strands. Most plasmids used for recombinant technology have recognition sequences for a number of restriction enzymes.