Using PCR for the in vitro mutagenesis of a protein coding sequence
The amino acid sequence across a region of interest in a protein is
Asn-Ser-Gly-Met-His-Pro-Gly-Lys-Leu-Ala-Ser-Trp-Phe-Val-Gly-Asn-Ser
The nucleotide sequence encoding this region begins and ends with anEcoRI site, making it easy to clone out the sequence and amplify it by polymerase chain reaction (PCR). Give the nucleotide sequence of this region. Suppose you wished to change the middle Ser residue to a Cys to study the effects of this change on the protein's activity. What would be the sequence of the mutant oligonucleotide you would use for PCR amplification?
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