NM494 - McrA +McrBC -EcoKR -EcoKM pop101 Strain NM494 has non functioning McrA,
ID: 140826 • Letter: N
Question
NM494 - McrA +McrBC -EcoKR -EcoKM pop101
Strain NM494 has non functioning McrA, functioning McrBC and non functioning EcoKR and EcoKM.
Y10 +McrA +McrBC +EcoKR +EcoKM
Strain y10 has functioning McrA, McrB, EcoKR, and EcoKM.
External Link: https://www.neb.com/tools-and-resources/usage-guidelines/mcra-mcrbc-and-ecoki-strain-phenotypes
. Suppose you isolate plasmid pUC19 from E. coli strain NM494 and then you transform the isolated plasmid into E. coli strain Y10. Will you be able to successfully transform this pUC19 sample into Y10? Explain why or why not, Use the strain phenotype information found here: https://www.neb.com/tools-and-resources/usage-guidelines/mcra-mcrbc-and-ecoki-strain- phenotypes. NOTE about the Table: In the Strain Phenotype Table, the "R" and "M" headings refer to the EcoKI restriction system, which is encoded by the hsdRMS genes. "R" refers to the hsdR gene, while "M" refers to the hsdM gene. "+" means the phenotype indicates normal gene activity (wild type), and""means that the gene activity is absent. (3 pts)Explanation / Answer
No, it is not possible to successfully transform the pUC19 plasmid from NM494 into Y10.
hsdR encodes for an endonuclease while hsdM encodes for methylase. The endonuclease cleaves unmethylated DNA.
NM494 strain is negative for hsdR and hsdM, this indicates that the plasmid is not methylated (since the plasmid is isolated from NM494). While Y10 is positive for both the genes. Transformation of an unmodified plasmid DNA into hsdR+ strains results in the fragmentation of the foreign gene. Therefore transformation of plasmid from NM494 into Y10 would result in its fragmentation.
The mcr genes are methycytosine restricting endonucleases, they participate in cleaving the methylcytosine in the DNA sequence.
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