The recognition sites of two restriction endonucleases are shownbelow. Which wou

Question

The recognition sites of two restriction endonucleases are shownbelow. Which would you use to excise the gene from the followingDNA fragment? Explain. You should write in the complementary DNAstrand before answering the question.

5'AAGGGCCCGAATTCGCGAATTCAGGGCCC---------gene---------TTGAATTC-3'

Recognition sites

EcoRI

5'-GAATTC-3'

3'-CTTAAG-5'

SmaI

5'-GGGCCC-3'

3'-CCCGGG-5'

5'AAGGGCCCGAATTCGCGAATTCAGGGCCC---------gene---------TTGAATTC-3'

Recognition sites

EcoRI

5'-GAATTC-3'

3'-CTTAAG-5'

SmaI

5'-GGGCCC-3'

3'-CCCGGG-5'

Explanation / Answer

The complementary strand is : 5'AAGGGCCCGAATTCGCGAATTCAGGGCCC---------gene---------TTGAATTC-3' 3'TTCCCGGGCTTAAGCGCTTAAGTCCCGGG------------------------AACTTAAG-5' The restriction enzyme to be used first isSmaI. The fragments it produces is: 5'AAGGGCCCGAATTCGCGAATTCAGGG         CCC---------gene---------TTGAATTC-3' 3'TTCCCGGGCTTAAGCGCTTAAGTCCC           GGG------------------------AACTTAAG-5' If we use the other enzyme EcoRI, we will get alarger fragment: 5'AAGGGCCCG                AATTCGCGAATTCAGGGCCC---------gene---------TTGAATTC-3' 3'TTCCCGGGCTTAA                  GCGCTTAAGTCCCGGG------------------------AACTTAAG-5' 5'AAGGGCCCGAATTCGCGAATTCAGGG         CCC---------gene---------TTGAATTC-3' 3'TTCCCGGGCTTAAGCGCTTAAGTCCC           GGG------------------------AACTTAAG-5' If we use the other enzyme EcoRI, we will get alarger fragment: 5'AAGGGCCCG                AATTCGCGAATTCAGGGCCC---------gene---------TTGAATTC-3' 3'TTCCCGGGCTTAA                  GCGCTTAAGTCCCGGG------------------------AACTTAAG-5' 5'AAGGGCCCG                AATTCGCGAATTCAGGGCCC---------gene---------TTGAATTC-3' 3'TTCCCGGGCTTAA                  GCGCTTAAGTCCCGGG------------------------AACTTAAG-5' After restriction by SmaI, we get the followingfragment with the gene: CCC---------gene---------TTGAATTC-3' GGG------------------------AACTTAAG-5'

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