Effect of pyk gene location on constructing dATP conversion E. coli strain

A dATP conversion E. coli strain could be constructed when both pyk and adk1 gene were expressed successfully. pyk gene encodes pyruvate kinase (PK) could be expressed, when inserted it before adk1 gene which encodes adenylate kinase (AK) in plasmid pET-pyk-adk1 after transform into E. coli and the recombinant could be used to convert dATP from dAMP. Another plasmid pET-adk1-pyk, which inserted pyk gene behind of adk1, the recombinant E. coli transformed with this plasmid could not convert dAMP into dATP, pyk gene cannot be translated in this recombinant. The different translation levels of pyk with gene location switching caused mainly by the different secondary structures formed by the 5’-untranslation regions and the gene sequence of its5’-terminal. The dATP product E. coli strain could be constructed when cloned pyk gene at an optimum location.