Cloning and expression of the Campylobacter jejuni lon gene detected by RNA arbitrarily primed PCR.
Fingerprinting of RNA by arbitrarily primed PCR was used to identify a heat-inducible gene in Campylobacter jejuni. Comparing RNA fingerprints from C. jejuni cells before and after 20 min of heat shock at 48 degrees C, a differentially amplified PCR product was identified which displayed a high degree of homology to bacterial lon genes. By screening C. jejuni genomic libraries, the entire lon gene was cloned and sequenced. It encodes a protein of 791 amino acids with a calculated molecular mass of 90.2 kDa. Alignment of the Lon amino acid sequence with that of other bacterial species revealed an overall identity of up to 56.6% (Helicobacter pylori). Northern and RNA dot blot experiments confirmed heat induction of the C. jejuni lon gene, revealing a maximum 6-8-fold increase in the level of specific mRNA.
Thies
F L
FL
Department of Neurology, University Würzburg, Germany.
Hartung
H P
HP
Giegerich
G
G
D049069:ATP-Dependent Proteases; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D016123:Campylobacter jejuni / Q000201:enzymology* / Q000235:genetics; D003001:Cloning, Molecular; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D018869:Heat-Shock Response; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids; D016133:Polymerase Chain Reaction / Q000379:methods*; D012329:RNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA; D012697:Serine Endopeptidases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D014158:Transcription, Genetic
D006360:Heat-Shock Proteins; D012329:RNA, Bacterial; D049069:ATP-Dependent Proteases; D012697:Serine Endopeptidases