Map The following table shows where different restriction endonucleases (restric

Question

Map The following table shows where different restriction endonucleases (restriction enzymes) cleave DNA. The abbreviation R represents the purines (adenine and guanine). The pyrimidines (cytosine, thymine, and uracil) are abbreviated as Y. The abbreviation W represents adenine or thymine Enzyme Target sequence Cleavage 5' G EcoRl EcORV Haelll Hindlll PpuMI 5' GAATTC 3' 3' CTTAAG 5 5' GATATC 3 3' CTATAG 5 5' GGCC 3' 3' CCGG 5 5' AAGCTT 3 3CTTAA AATTC 3 G 5' 5' GAT ATC 3 3' CTA TAG 5 5' GG CC 3 ?'CC GG5 5' A 3' TTCGA 5' RG ?' YCCWG AGCTT 3 3. TTCGAA 5 5' RGGWCCY 3 A 5' GWCCY 3 GR 5 3' YCCWGGR 5 Which restriction endonuclease(s) would cleave a DNA molecule at the following sequences? a) 5' GCCAATTGAAGCTT 3 b) 5 TCCAAGCTTGAATTC 3 Haelll EcoRV Hindll EcoRI ? EcoRI ? HindIII EcoRV ? Haelll continued below... Plasmid DNA and a gene of interest are cut with the en the sticky end of the gene in the 5' to 3' direction zyme PpuMI. Write a possible sequence of bases for 5' 3'

Explanation / Answer

a) 5' GCCAATTGAAGCTT 3'

This will be cleaved by HindIII as it has the restriction site AAGCTT

GCCAATTGA / AGCTT

b) 5' TCCAAGCTTGAATTC 3'

This will be cleaved by two endonucleases EcoRI and HindIII

EcoRI ----- TCCAAGCTTG / AATTC

HindIII ------ TCCA / AGCTTGAATTC

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