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To produce a recombinant plasmid and the foreign DNA are cut with a different restriction enzyme?
Fasle.
Which Restriction enzyme are studied in Recombinant DNA Technology?
It's not the restriction enzymes that are studied, its the DNA. The enzyme cuts or "restricts" the DNA strand at a known sequence of nucleotides. Different enzyme, different sequence. For a Biomanufacturing application, where we want to insert foreign DNA, the gene of interest is cut and spliced with a restriction enzyme into a recombinant plasmid, transformed into a bacteria, and sent merrily on it's way to make Insulin, or whatever. With an unknown piece of DNA (a functional gene that makes a protein of interest or is being studied), the plasmid has "restriction sites" or nucleotide sequences, for several restriction enzymes, all of which I have mapped out. The unknown piece of DNA is cut at each end by a single restriction enzyme and inserted into the plasmid, which gives me some landmarks. I insert the plasmid into a bacteria, grow a culture so the bacteria makes many millions of copies of the plasmid, extract the plasmid, and run an experiment called a restriction digest. The restriction digests are a series of reaction with single enzyme and combinations of two and three enzymes, all cutting the plasmid at different nucleotide sequences. Then I run an agarose gel electrophoresis, which separates all the different pieces of DNA by size, and do an analysis called a Restriction Map. This counts the DNA fragments and their sizes, which enzyme and combination of enzymes produced which sizes and how many fragments, which enzyme cuts where, which cuts were definitely in the known part of the plasmid, which were probably in the unknown DNA, adding up nucleotide sequence numbers to make sure different mapping guesses agree, etcetera, etcetera, and so forth. Until at last, a map of the size and restriction sites of the unknown DNA insert into the known plasmid vector is deduced. This used to be done by hand, but there are computer programs that do it now. This is Research, the Technology is down the line a few steps when the gene has been characterized, the protein produced has been characterized, the trials are done, and the restriction enzyme to insert the gene into the bacteria for Bioman has been established
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)
When you run a gel of a plasmid that has been digested with a single enzyme for which there is a single restriction site on the plasmid why do you sometimes get more then one band?
There are two possible reasons, and it may help enrich your digest concentrations by looking into these two possible solutions: 1. Incomplete digestion. The migration of an untreated plasmid travels faster that linearized DNA. The apparent band size of a circular DNA may be as much as 500bp smaller than its linearized counterpart. 2. Perhaps the agar wasn't fully melted when the gel was prepared. This can cause resolution problems, and sometimes a smear would show up. 3. The enzyme used is one of the more uncommon enzymes (noted by the volume the enzyme came in), and so these may be more picky at its turnover rate, leading to partial digestions. Most of the time a restriction enzyme does its job really well if it was allowed to digest 1-2ug of DNA for at least 2 hours for NEB brand of enzymes. If it was just a diagnostic restriction digest, then you can predict what those bands would look like if it was a partial digest. Good luck.
Why is it important to use the same restriction enzyme for both cells in recombinant DNA?
Restriction enzymes are endonucleases that digest the DNA at a sequence specific site. Hind III for example cut between two As in the sequence AAGCTT in the both strand forming a sticky end. If you use this enzyme to cut in your vector DNA, you have to use the same enzyme in the insert DNA so as they can ligate by DNA ligation. This is the important use of same restriction enzyme in cloning.
To produce a recombinant plasmid and the foreign DNA are cut with a different restriction enzyme?
Fasle.
Which Restriction enzyme are studied in Recombinant DNA Technology?
It's not the restriction enzymes that are studied, its the DNA. The enzyme cuts or "restricts" the DNA strand at a known sequence of nucleotides. Different enzyme, different sequence. For a Biomanufacturing application, where we want to insert foreign DNA, the gene of interest is cut and spliced with a restriction enzyme into a recombinant plasmid, transformed into a bacteria, and sent merrily on it's way to make Insulin, or whatever. With an unknown piece of DNA (a functional gene that makes a protein of interest or is being studied), the plasmid has "restriction sites" or nucleotide sequences, for several restriction enzymes, all of which I have mapped out. The unknown piece of DNA is cut at each end by a single restriction enzyme and inserted into the plasmid, which gives me some landmarks. I insert the plasmid into a bacteria, grow a culture so the bacteria makes many millions of copies of the plasmid, extract the plasmid, and run an experiment called a restriction digest. The restriction digests are a series of reaction with single enzyme and combinations of two and three enzymes, all cutting the plasmid at different nucleotide sequences. Then I run an agarose gel electrophoresis, which separates all the different pieces of DNA by size, and do an analysis called a Restriction Map. This counts the DNA fragments and their sizes, which enzyme and combination of enzymes produced which sizes and how many fragments, which enzyme cuts where, which cuts were definitely in the known part of the plasmid, which were probably in the unknown DNA, adding up nucleotide sequence numbers to make sure different mapping guesses agree, etcetera, etcetera, and so forth. Until at last, a map of the size and restriction sites of the unknown DNA insert into the known plasmid vector is deduced. This used to be done by hand, but there are computer programs that do it now. This is Research, the Technology is down the line a few steps when the gene has been characterized, the protein produced has been characterized, the trials are done, and the restriction enzyme to insert the gene into the bacteria for Bioman has been established
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)
If you took a linear piece of DNA and cut it with the restriction enzyme EcoRI and it had three restriction sites for EcoRI, how many fragments would you produce What if you had a circular piece of DNA?
If the plasmid has 3 recognition sequences for a given restriction endonuclease, then 4 linear DNA fragments are obtained because, if the DNA is linear then the number of fragments obtained is (N+1) whereas if the DNA is circular then the number of fragments obtained will be N for N recognition sequences for the given restriction endonuclease in a plasmid.
When you run a gel of a plasmid that has been digested with a single enzyme for which there is a single restriction site on the plasmid why do you sometimes get more then one band?
There are two possible reasons, and it may help enrich your digest concentrations by looking into these two possible solutions: 1. Incomplete digestion. The migration of an untreated plasmid travels faster that linearized DNA. The apparent band size of a circular DNA may be as much as 500bp smaller than its linearized counterpart. 2. Perhaps the agar wasn't fully melted when the gel was prepared. This can cause resolution problems, and sometimes a smear would show up. 3. The enzyme used is one of the more uncommon enzymes (noted by the volume the enzyme came in), and so these may be more picky at its turnover rate, leading to partial digestions. Most of the time a restriction enzyme does its job really well if it was allowed to digest 1-2ug of DNA for at least 2 hours for NEB brand of enzymes. If it was just a diagnostic restriction digest, then you can predict what those bands would look like if it was a partial digest. Good luck.
Why is it important to use the same restriction enzyme for both cells in recombinant DNA?
Restriction enzymes are endonucleases that digest the DNA at a sequence specific site. Hind III for example cut between two As in the sequence AAGCTT in the both strand forming a sticky end. If you use this enzyme to cut in your vector DNA, you have to use the same enzyme in the insert DNA so as they can ligate by DNA ligation. This is the important use of same restriction enzyme in cloning.
An enzyme that cuts double-stranded DNA at specific nucleotide sequences?
Such an enzyme is called a restriction endonuclease
When circular plasmid DNA is cut open at a single site by a restriction enzyme the resulting piece is?
You should end up with a linear piece of DNA the length of the entire plasmid. I don't know if there is a specific name for it. CALLED A : a linear strand
What determines how DNA will be cut by a restriction enzyme?
Restriction enzymes cut DNA at specific sites called restriction sites. These restriction sites are typically 6 - 8 nucleotides in length and have a defined set of nucleotide bases. For example, the restriction enzyme Eco R1 cuts at the site: AGGTTC. Therefore, if the target DNA contains the above sequence, Eco R1 is able to cut it within the restriction site. Hence, by looking into the target site and which restriction enzymes are being used, on can make an accurate estimate of where the target DNA will be cut
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.
The restriction enzyme used in constructing hybrid molecules of certain gene sequences and plasmid DNA acts by?
opening DNA molecules at specific sites,leaving sticky ends exposed. find me on kik one1992
What enzyme cuts and seals DNA?
HaeIIIrestriction enzymes
