What process is the researcher intending to detect with the use of these restric
ID: 259112 • Letter: W
Question
What process is the researcher intending to detect with the use of these restriction enzymes?
Select all that apply.
a. mutations that create new SmaI recognition sites
b. mutations that disrupt PvuII restriction sites
c. mutations that create new PvuII restriction sites
d. mutations that disrupt SmaI restriction sites
Can anyone help me with this question? Please and thank you in advance!
A geneticist searching for mutations uses the restriction endonucleases SmaI and PvuII to search for mutations that eliminate restriction sites. SmaI will not cleave DNA with CpG methylation. It cleaves DNA at the restriction digestion sequence 3'-GGG CCC-5, PvulI is not sensitive to CpG methylation. It cleaves DNA at the restriction sequence 5'-CAG CTG-3 3'-GTC GAC-5,Explanation / Answer
According to me all four options can be occured. As here restriction site is CpG containing sequence so methylation can be happended where CpG sequence is present. Random mutation can either create or disrupt restriction as well as recognition sites. Both these enzymes are belong to type II enzyme and thus their restriction sites are at or near recognition sites. If new CpG site is created after mutation then methylation can be happened in that site. Above sequences both contain CpG sequence but SmaI is not able to cleave DNA with CpG methylation where as PvuII is not sensitive to CpG methylation and can cleve DNA with CpG methylation. So one possibility is that mutation can alter the restriction site within the DNA and thus the restriction enzymes will be prevented to recognize that site as well as to cut during a restriction digest. And another possibility is new recognition sites will be created cleavage on the sequence will be occured. Based on methylation sensitivity property, if mutation disrupt the restriction site then no cleavege product will be formed and if new recognition site is created then more cleavage products will be obtained besides cleavage products obtained from normal restriction site cut.
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