Question

Plasmid DNA and a gene of interest are cut with the enzyme PpuMI. Write a possible sequence of bases for the sticky end of the gene in the 5' to 3' direction.

Answer 1

Answer:

5' RG        GWCCY 3'

3' YCCWG        GR 5'

Explanation:

The enzyme PpuMI is a restriction endonuclease enzyme, it has a specific recognition site where it cut the DNA. The source of the enzyme is from ​​an E. coli strain that carries the PpuMI gene from Pseudomonas putida (R. Morgan).

The enzyme PpuMI recognizes specific sequence with palindrome arrangement. It target the sequence 5' RGGWCCY 3'

target Sequence: 5' RGGWCCY 3'

                            3' YCCWGGR 5'

The enzyme cleavage point is at:

5' RG^GWCCY 3'

3' YCCWG^GR 5'

The product of the cleavage will give a sticky end Cleavage:

5' RG        GWCCY 3'

3' YCCWG        GR 5'

Note: R stands for purines (adenine and guanine). Y stands for pyrimidines (cytosine, thymine, and uracil). And W represents adenine or thymine.