[Construction of a Pseudomonas aeruginosa random promoter library and identification of novel iron-regulated genes by high-throughput library screening].
OBJECTIVE
Iron is an essential element for bacterial survival and pathogenesis. Using a random promoter library we screened for iron-responsive genes in P. aeruginosa genome.


METHODS
Plasmid pMS402 containing a promoterless luminescence reporter, LuxCDABE, was used as the vector to construct a P. aeruginosa random promoter library with a size of about 5700 clones. It enabled real-time, in vivo high-throughput gene expression profiling at genomic scale.


RESULTS
Two previously reported iron-regulated genes were identified, and ten new iron-regulated genes were uncovered. The genes encoding dihydrolipoamide acetyltransferase, phosphogluconate dehydratase and Fe (III) dicitrate transporter were found to be iron-regulated together with four function-unknown genes and three putative protein encoding genes.


CONCLUSIONS
These results provide a basis for elucidating the complex iron regulation network in P. aeruginosa and help our understanding of the roles of iron in bacterial physiology and pathogenesis. The random promoter library system also offers a useful tool in bacterial genomic studies.
Wang
Yan
Y
Key Laboratory of Resources Biology and Biotechnology in Western China, Ministry of Education, Faculty of Life Sciences, Northwest University, Xi'an, 710069, China. 900smile@163.com
Wei
Jinhua
J
Shen
Lixin
L
Duan
Kangmin
K
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D005798:Genes, Bacterial / Q000235:genetics*; D016680:Genome, Bacterial / Q000235:genetics; D015698:Genomic Library; D007501:Iron / Q000494:pharmacology*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011550:Pseudomonas aeruginosa / Q000166:cytology / Q000187:drug effects / Q000235:genetics*
D001426:Bacterial Proteins; D007501:Iron