Nucleotide Sequence Determination, Subcloning, Expression and Characterization of the xy1LT Region of the Pseudomonas putida TOL Plasmid pDK1

• Main Author: Baker, Ronald F. (Ronald Fredrick)
• Other Authors: Benjamin, Robert C.
• Format: Others
• Language: English
• Published: University of North Texas 1992
• Subjects: Pseudomonas putida.; Plasmids > Genetics.; nucleotide sequence; subcloning
• Online Access: https://digital.library.unt.edu/ark:/67531/metadc278316/

The complete nucleotide sequence of the region encoding the DHCDH function of the pDK1 lower operon was determined. DNA analysis has shown the presence of two open reading frames, one gene consisting of 777 nucleotides encoding a polypeptide of 27.85 kDa and another gene of 303 nucleotides encoding a polypeptide of 11.13 kDa. The results of enzymatic expression studies suggest that DHCDH activity is associated only with xy1L. However although the addition of xy1T cell-free extracts to xy1L cell-free extracts does not produce an increase in DHCDH activity, subclones carrying both xy1L and xy1T exhibit 300- 400% more DHCDH activity than subclones carrying only xy1L.