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How does bacterial cell protect its own DNA from restriction enzymes?
In order to protect the bacterial genomic DNA from its own restriction enzymes, bacterial cells employ a system, wherein methyl transferases methylate certain bases on the DNA sequence, making them unrecognizable to the restriction enzymes.Each restriction enzyme has a methylase associated with it on the chromosome. This methylase puts methyl groups on the host DNA, and the restriction enzyme doesn't recognize its recognition sequence when it is so methlyated. The host DNA is thus protected from the actions of its own restriction enzyme.Incoming (foreign) DNA is unlikely to be protected (methylated) in the same manner, thus this invading DNA is digested by the hosts restriction enzyme(s).When working in cloning experiments, the principle is the same -- DNA to be digested is carried by a plasmid in a host that does not methylate DNA in the pattern that would cause the restriction enzyme to see it as protected, thus it is cut. DNA generated by PCR is similarly unmethylated, and is therefore also digested.Some enzymes won't cut DNA isolated from dam+ or dcm+ hosts (two common bacterial methylases), thus one must know the genotype of the host cloning strain if using a restriction enzyme whose action is blocked by dam ordcmmethylation.
What is the restriction site of the restriction enzyme TaqI?
TaqI's restriction site is:TCGAAGCT
What is the function of restriction enzymes in the process of DNA recombination?
First, a specific enzyme is needed to cut the DNA from the donor genes at a specific site. This enzyme is called a restriction enzyme.The enzyme is used to cut out a piece of DNA that contains one or more desired genes from the donor's DNA. Next, a vector is needed to receive the donor DNA. Most frequently, a naturally occurring circular piece of bacterial DNA, called a plasmid, is used for this purpose. Finally, an enzyme is used to "stitch" the donor DNA into the plasmid vector. This enzyme is called ligase, and it creates permanent bonds between the donor DNA and the plasmid DNA. The result is that the donor DNA is incorporated into the bacterial plasmid, forming the recombinant DNA (rDNA)
Why did you cut both segments of DNA with the same restriction enzyme?
You use the same enzyme inn order to get the same restriction and binding sites.
What is the restriction site of the restriction enzyme Hae III?
The restriction site of Hae III is GGCC. It cuts between the G and the C. This produces blunt ends.
Which enzyme is Known as Genetical or Genetic Knife?
restriction endonuclase enzyme (made in bacterial plasmids)
How can a bacterium produce restriction enzymes that do not cleave its DNA?
In order to protect the bacterial genomic DNA from its own restriction enzymes, bacterial cells employ a system, wherein methyl transferases methylate certain bases on the DNA sequence, making them unrecognizable to the restriction enzymes.Each restriction enzyme has a methylase associated with it on the chromosome. This methylase puts methyl groups on the host DNA, and the restriction enzyme doesn't recognize its recognition sequence when it is so methlyated. The host DNA is thus protected from the actions of its own restriction enzyme.Incoming (foreign) DNA is unlikely to be protected (methylated) in the same manner, thus this invading DNA is digested by the hosts restriction enzyme(s).When working in cloning experiments, the principle is the same -- DNA to be digested is carried by a plasmid in a host that does not methylate DNA in the pattern that would cause the restriction enzyme to see it as protected, thus it is cut. DNA generated by PCR is similarly unmethylated, and is therefore also digested.Some enzymes won't cut DNA isolated from dam+ or dcm+ hosts (two common bacterial methylases), thus one must know the genotype of the host cloning strain if using a restriction enzyme whose action is blocked by dam ordcmmethylation.
Where restriction enzymes come from?
Restriction enzyme, also called restriction endonuclease, a protein produced by bacteria that cleaves DNA at specific sites along the molecule. In the bacterial cell, restriction enzymes cleave foreign DNA, thus eliminating infecting organisms.
An enzyme that cuts double-stranded DNA at specific nucleotide sequences?
Such an enzyme is called a restriction endonuclease
How does bacterial cell protect its own DNA from restriction enzymes?
In order to protect the bacterial genomic DNA from its own restriction enzymes, bacterial cells employ a system, wherein methyl transferases methylate certain bases on the DNA sequence, making them unrecognizable to the restriction enzymes.Each restriction enzyme has a methylase associated with it on the chromosome. This methylase puts methyl groups on the host DNA, and the restriction enzyme doesn't recognize its recognition sequence when it is so methlyated. The host DNA is thus protected from the actions of its own restriction enzyme.Incoming (foreign) DNA is unlikely to be protected (methylated) in the same manner, thus this invading DNA is digested by the hosts restriction enzyme(s).When working in cloning experiments, the principle is the same -- DNA to be digested is carried by a plasmid in a host that does not methylate DNA in the pattern that would cause the restriction enzyme to see it as protected, thus it is cut. DNA generated by PCR is similarly unmethylated, and is therefore also digested.Some enzymes won't cut DNA isolated from dam+ or dcm+ hosts (two common bacterial methylases), thus one must know the genotype of the host cloning strain if using a restriction enzyme whose action is blocked by dam ordcmmethylation.
Why restriction enzyme cannot cut its own DNA?
Restriction enzymes are produced by bacteria to help destroy foreign, invading DNA, such as the DNA of bacteriophage (a virus that infects bacterial cells). Every restriction enzyme comes with a methylase enzyme, or more specifically, a DNA methyltransferase. The methylase enzyme methylates (adds a methyl group) to the restriction endonuclease site on the cell's own DNA, which protects the sites from the restriction enzyme so that it does not degrade its own DNA.
Which restriction enzyme did you use to cut the DNA?
The restriction enzyme used to cut the DNA was EcoRI.
How do restriction sites and a restriction map relate?
Restriction sites are specific DNA sequences recognized and cleaved by restriction enzymes, while a restriction map shows the locations of these sites on a DNA molecule. A restriction map provides information on the order and spacing of restriction sites along a DNA sequence, helping to identify the size and organization of DNA fragments generated by restriction enzyme cleavage.
How can bacterium produce restriction enzymes that do not cleave it's DNA?
Restriction enzymes cleave DNA at a particular recognition site -- a particular sequence of nucleotides. You can imagine the following scenarios:1. The bacterial chromosome does not contain the recognition sequence2. The bacterial chromosome contains the recognition sequence, but that particular part of the DNA is either supercoiled to keep the restriction enzyme from finding the sequence, or it's single stranded as when being replicated or transcribed.3. The bacterial chromosome contains the recognition sequence, but that particular part of the DNA is methylated or modified in some other way which prevents the restriction enzyme from attaching.
Hi does any one know if I want to put some restriction enzyme close to cDNA should I put some nucleotide befor this restriction enzyme?
Restriction enzyme cuts DNA strand at specific locations Restriction enzyme cuts DNA strand at specific locations
Restriction enzyme digestion?
Yes?
Where does the restriction enzyme EcoRI cut in a DNA sequence?
The restriction enzyme EcoRI cuts DNA at a specific sequence of bases, which is GAATTC.
