OBJECTIVE To identify and construct and overexpress a heat resistant and high active Pyrimidine Nucleotide Phosphorylase(PyNPase) isolated form soil.METHODS PyNPase gene cloned with the degenerate PCR method,based on which a pair of special primers were designed according to PyNPase nucleotide sequence.And Nco I,Xba I recognising sites were introduced into 5'-and 3'-primers.An expression vector containing a(His-tag)6 was constructed by inserting pynp gene into pBAD/gIII A.Then transferred into E.coli TOP10F...
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