Premium
Differential gene expression in response to copper in Acidithiobacillus ferrooxidans analyzed by RNA arbitrarily primed polymerase chain reaction
Author(s) -
Paulino Luciana Campos,
de Mello Maricilda P.,
Ottoboni Laura M. M.
Publication year - 2002
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200202)23:4<520::aid-elps520>3.0.co;2-r
Subject(s) - gene , caulobacter crescentus , biology , operon , polymerase chain reaction , microbiology and biotechnology , gene product , gene expression , biochemistry , escherichia coli , bacterial protein
Acidithiobacillus ferrooxidans is a chemoautotrophic bacterium that plays an important role in metal bioleaching processes. Despite the high level of tolerance to heavy metals shown by A. ferrooxidans , the genetic basis of copper resistance in this species remains unknown. We investigated the gene expression in response to copper in A. ferrooxidans LR using RNA arbitrarily primed polymerase chain reaction (RAP‐PCR). One hundred and four differentially expressed genes were identified using eight arbitrary primers. Differential gene expression was confirmed by DNA slot blot hybridization, and approximately 70% of the RAP‐PCR products were positive. The RAP‐PCR products that presented the highest levels of induction or repression were cloned, sequenced and the sequences were compared with those in databases using the BLAST search algorithm. Seventeen sequences were obtained. The RAP‐PCR product with the highest induction ratio showed similarity with the A. ferrooxidans cytochrome c . A high similarity with the thiamin biosynthesis gene thi C from Caulobacter crescentus was observed for another RAP‐PCR product induced by copper. An RAP‐PCR product repressed by copper showed significant similarity with the carboxysome operon that includes the ribulose‐1,5‐bisphosphate carboxylase/oxygenase complex from A. ferrooxidans and another copper‐repressed product was significantly similar to the XyIN outer membrane protein from Pseudomonas putida . Finally, RAP‐PCR products of unknown similarities were also present.