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Evaluation · Q19

Q.You are working in a biotechnology lab with a becterium namely E.coli. How will you cut the nucleotide sequence? explain it.

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Step 1. Identify the right tool. Cutting a DNA/nucleotide sequence at a specific point is exactly the job of a restriction enzyme (restriction endonuclease), described in the chapter as a molecular scissor that cleaves DNA into fragments at or near specific recognition sites.

Step 2. Choose an appropriate enzyme for E. coli work. Since the scenario specifically involves E. coli, EcoRI - itself originally isolated from Escherichia coli - is a natural, directly relevant choice; it recognises the palindromic sequence GAATTC and cuts it in a staggered way to leave sticky ends.

Step 3. Explain the mechanism. The enzyme scans the DNA for its specific recognition sequence and cleaves the phosphodiester backbone at (or near) that site on both strands, producing defined DNA fragments with either blunt or sticky ends depending on the enzyme used. …

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