Expression of the flgFG operon of Campylobacter jejuni in Escherichia coli yields an extra fusion protein.

Abstract

Two Campylobacter jejuni genes with homology to the Escherichia coli flgF and flgG genes encoding two of the basal body rod proteins were isolated, and the nucleotide sequence was determined and analyzed. These two C. jejuni genes were shown, by Northern hybridization analysis, to function as a single operon (flgFG). Two transcriptional start sites were detected upstream of flgF, corresponding to the two RNA transcripts detected in the Northern blot. Western blot immunoassays using anti-FlgF and anti-FlgG antibodies demonstrated the synthesis of FlgF and FlgG proteins in C. jejuni and in Escherichia coli containing the C. jejuni flgF and flgG genes. Maxicell analysis and Western immunoblots using anti-FlgF antibodies to probe flgFG-encoded proteins in E. coli revealed the presence of a protein with a molecular mass of approximately the combined mass of the FlgF and FlgG proteins. Anti-FlgF antibodies detected in C. jejuni cell extracts the native FlgF protein and also a higher-molecular-weight protein that is likely encoded by the flgF and part of the flgG sequences.