ISSN:
1573-5028
Keywords:
recombinant
;
phosphoenolpyruvate carboxylase
;
C4 plant
;
cDNA
;
transformation
;
Escherichia coli
;
protein phosphorylation
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract Phosphoenolpyruvate carboxylase (PEPC)-deficient mutants ofEscherichia coli have been complemented with a plasmid bearing a full-length cDNA encoding the C4-type form ofSorghum leaf PEPC. Transformed cells grew on minimal medium. Two clones were selected which produce a functional and full-sized enzyme protein as determined by activity assays, immunochemical behavior and SDS-PAGE. In addition, regulatory phosphorylation of immunopurified recombinant PEPC was observed when the enzyme was incubated with a partially purified plant PEPC kinase. These results establish thatE. coli cells produce a genuine, phosphate-free, higher-plant PEPC. Application of immunoadsorbtion chromatography to bacterial extracts makes it possible to prepare highly pure protein available for biochemical studies.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00036808
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