Q4) What’s the evolutionary purpose of restriction enzymes? Why is the bacterial DNA not harmed in this process? Q5) Restriction enzymes look for a very particular KIND OF sequence. What is that called. Give me an example of one.
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Q4) What’s the evolutionary purpose of restriction enzymes? Why is the bacterial DNA not harmed in this process?
Q5) Restriction enzymes look for a very particular KIND OF sequence. What is that called. Give me an example of one.
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- Q5) Restriction enzymes look for a very particular KIND OF sequence. What is that called. Give me an example of one.As you know, restriction enzymes evolved in different bacterial species independently. The adaptive significance of having a restriction enzyme is that the bacterium has the ability to cut the injected viral DNA into small segments. This destruction of viral DNA prevents the virus from taking over the bacterial cell and killing the cell. What is one benefit of using a restriction enzyme with staggered ends (such as EcoRI) to cut both the DNA insert and the plasmid? Which types of cut sites (staggered with “sticky ends” or blunt ends) are most useful in cloning DNA? Would you expect restriction enzymes in different bacteria genera (Streptococcus, Lactobacter, Escherichia) to have the same recognition sites (DNA sequences). Why or why not?Which of the following is true about restriction endonucleases?a) Type I and II requires ATP to move along DNAb) Type I, II and III requires ATP to move along DNAc) Type II requires no ATP and cleaves DNA within recognition sequenced) Type II requires ATP and cleaves DNA within recognition sequence
- A small DNA molecule was cleaved with several different restriction nucleases, and the size of each fragment was determined by gel electrophoresis.The following data were obtained. (a) Is the original molecule linear or circular?(b) Draw a map of restriction sites (showing distances between sites) that isconsistent with the data given.(c) How many additional maps are compatible with the data?(d) What would have to be done to locate the cleavage sites unambiguouslywith respect to each other?Which of the following is necessary for a PCR reaction to proceed? a) the sequence of the ends of the DNA to be amplified must be known. b) the sequence of restriction endonuclease recognition sites in the DNA to be amplified and in the plasmid, where the amplified DNA fragment will be cloned must be known. c) The complete sequence of the DNA to be amplified must be known. d) The sequence of restriction endonuclease recognition sites in the DNA to be amplified must be knownA) For this DNA fragment (from 5' to 3') "TGAATTCCCGGGTTCCGGGAATTCGCGCGAATTCCCGGTATA", what is its complementary strand B) What are the products when the DNA with the above sequence is incubated with the restriction enzyme EcoRI C) What are the products when the DNA with the above sequence is incubated with the restriction enzyme Mspl D) Draw the first two (2) base pairings of the DNA molecule from the 5' end and label all key elements of the molecule including the bonds involved
- What sequence do the restriction enzymes used in the lab recognize? How do they cut? And how would these different cuts effect cloning? (i.e. compare and contrast how overhang and blunt cuts will differ with cloning.) What organisms were the restriction enzymes used in this lab derived from? Why are restriction enzymes important for the organism that makes them? What is the purpose of multiple cloning sites on pUC19? What size were each of your plasmid fragments?Restriction sites are palindromic; that is, they read the same in the5' to 3' direction on each strand of DNA. What is the advantage ofhaving restriction sites organized this way?Restriction endonuclease and ligase are two types of enzymes used in the process of genetic engineering, i.e., the manipulation of genes. The restriction endonuclease differs from ligase in that it breaks the DNA at ends, while ligase causes the breaks in DNA from interior joins the fragments of DNA, while ligase breaks the DNA into fragments breaks the DNA at specific points, while the ligase joins the fragments of DNA breaks the DNA apart at each nucleotide, while ligase use the pieces to translate
- Q6) Why are ‘sticky ends’ useful biotechnologically? Q7) Why is this enzyme called a restriction endonuclease? How is it different from an exonuclease?In making recombinant DNA molecules that combine restriction fragments from different organisms, researchers usually prefer restriction enzymes like BamHI or HindIII that generate fragments with “sticky ends” (ends with overhangs) rather than enzymes like HpaI or SmaI (Table 12.1) that generate fragments with “blunt ends” (ends without overhangs). Can you think of a reason for this preference?a) what are restriction enzymes? b) What is the main function of restriction enzymes in nature? c) Compare and contrast the these enzymes in nature and in scientific research.