Isolation of the Rhodopseudomonas sphaeroides form I ribulose 1,5-bisphosphate carboxylase/oxygenase large and small subunit genes and expression of the active hexadecameric enzyme in Escherichia coli.
A library of cloned Rhodopseudomonas sphaeroides DNA was screened by colony hybridization for form I ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBPC/O) sequences using heterologous RuBPC/O probes. A recombinant plasmid was identified that hybridized to both the Anacystis nidulans and the R. sphaeroides form II RuBPC/O genes. Subcloning of a hybridizing 4-kb SmaI fragment allowed expression of active enzyme in Escherichia coli that was identical to form I RuBPC/O based on polyacrylamide gel electrophoresis and Western immunoblot analysis.
D004262:DNA Restriction Enzymes; D004926:Escherichia coli / Q000235:genetics*; D005796:Genes; D005798:Genes, Bacterial; D005838:Genotype; D046911:Macromolecular Substances; D009693:Nucleic Acid Hybridization; D010641:Phenotype; D010957:Plasmids; D012242:Rhodobacter sphaeroides / Q000201:enzymology / Q000235:genetics*; D012273:Ribulose-Bisphosphate Carboxylase / Q000235:genetics*
D046911:Macromolecular Substances; D004262:DNA Restriction Enzymes; D012273:Ribulose-Bisphosphate Carboxylase