The cloning of cytosine deaminase gene of Escherichia coli and the construction of eukaryotic expression vector

Objective To clone cytosine deaminase(CD) gene of Escherichia coli and to construct its eukaryotic expression vector Methods A pair of primers were designed and synthesized based on CD gene sequence in GenBank And CD gene was cloned from Escherichia coli genome by PCR amplification The PCR products were cloned into a eukaryotic plasmid pcDNA3 1 to construct the recombination expression vector which was controlled by CMV promoter The recombinant plasmid was analyzed and identified by PCR, restriction digest and sequencing Results The CD gene was cloned and eukarytoic expression vector (pcDNA3 1 CD) was constructed It was demonstrated that CD gene was properly inserted into the vector and the sequence was confirmed with restriction digest, PCR amplification and sequencing Conclusion The eukaryotic expression vector with CD gene (pcDNA3 1 CD) was successfully constructed