Open AccessCloning and expression of the Campylobacter jejuni lon gene detected by RNA arbitrarily primed PCR
Author(s) -
Thies Frank L.,
Hartung HansPeter,
Giegerich Gerhard
Publication year - 1998
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1998.tb13165.x
Subject(s) - cloning (programming) , campylobacter jejuni , biology , gene , campylobacter , gene expression , microbiology and biotechnology , genetics , bacteria , computer science , programming language
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°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.