How are restriction enzymes used in the laboratory?

Answer 1

Restriction enzymes are used to cut up DNA into fragments with 'sticky ends'. It allows for the gene of interest to be isolated. A plasmid can then also be cut with the same restriction enzyme and the sticky ends are spliced together with DNA ligase. The recombinant plasmid can then be put into new host cells via a variety of methods.

Answer 2

Restriction enzymes are often sourced from bacteria. They cut DNA at specific sequences and so can be used to create fragments of DNA from whole strands. The characteristic pattern of these fragments for different restriction enzymes is used in genetic finger printing of people and bacteria.

Answer 3

Plasmids, circular double stranded DNA molecules, are used as a shuttle to transport a gene of interest into a foreign host cell for a number of reasons. These reasons include either maintenance of the plasmid within the cytoplasm, or integration of the foreign DNA into the nuclear DNA of the host cell. If the plasmid is maintained in the cytoplasm this often to express a heterologous protein, and has significant application in the commercial sector where bioreactors generate substantial income from production of proteins/enzymes for a multitude of applications. Thus an organism has been genetically engineered with a specialized protein expression vector (plasmid) to express a foreign protein. Expression of foreign proteins are also essential for protein studies where a purified form of a protein is required to assess and characterize its structure and function. Another application of plasmid transfer to the cytoplasm of a bacterial cell is present in increasing the copy number (amount of plasmids) of the DNA. When large numbers of plasmids have been generated the cells can be lysed and the plasmids isolated for further use and additional transformation experiments. Construction of DNA libraries also require the use of plasmids to have genomic DNA of the organism in question inserted within them and maintained for screening of genes of interest. Some unique vectors based on the principles of Ti-plasmid of Agrobacterium species can transfer its DNA into the genome of the host cell by a process of non-homologous end joining that is not sequence specific. This insertion does tend to be localized to certain areas of the genome, and is a popular transformation technique of dicot plant species. Getting back to restriction enzymes, and straining their importance as genetic engineering would not be possible without their existence. The Restriction enzymes can be compared to molecular scissors that excise DNA sequences from a larger DNA segment. They are naturally occurring enzymes present in all living organisms and a large number have been made commercially available for industrial or research purposes. The restriction enzyme recognizes a specific sequence within a DNA sequence known as a restriction site, and can make a single stranded or double stranded cut depending on the restriction enzyme used. The cut if double stranded generates two possible scenarios, either a blunt end or a sticky-end overhang. Different restriction enzymes recognize different restriction sites and provide an opportunity to isolate specific regions of DNA when sequence information is available. After isolation of a specific DNA sequence, primers are designed, and the sequence is amplified by means of PCR. In parallel a mixture is prepared containing a plasmid with a multiple cloning site containing numerous restriction sites. The plasmid is then incubated with one or 2 restriction enzymes depending on the sequence of the DNA to be inserted, and allows for orientation of the gene to ensure expression under a valid promoter in the plasmid. Natural base pairing of DNA takes place and complementary sticky ends are ligated with DNA ligase to incorporate the gene of interest into the plasmid.

Answer 4

Restriction enzymes are used to isolate genes of interest or to insert genes into different genomes (recombination). Essentially, they allow geneticists to modify the genome of organisms - which is why we have genetically-modified foods.

Answer 5

restriction enzymes cut the DNA at specific sites called restriction sites. These restriction sites are 6 - 8 base pairs long and occur at different frequencies on the DNA

Answer 6

The restriction enzyme is used to "cut" the insulin gene and place it in the plasmid of the bacterium :) hope this helps.

Answer 7

restrication enzyme in rdt used as a scisseors i.e it use for cutting the the DNA into small fragments.

Answer 8

Restriction enzymes, ligases and reverse transcriptase are used in gene transfer.

Answer 9

I really don't know... That's a hard one😕

Answer 10

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