ArticleApplied and environmental microbiology2006
An integrated approach to functional genomics: construction of a novel reporter gene fusion library for Sinorhizobium meliloti.
Article in Applied and environmental microbiology, 2006. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 38 papers.
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Who cites it
38 citing papers in PubMed, 78 citations in OpenAlex.
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- A minimized symbiotic gene set from the 1.68 Mb pSymB chromid of Sinorhizobium meliloti reveals auxiliary symbiotic loci.BMC biology · 2025Article
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- Symbiotic and Nonsymbiotic Members of the Genus Ensifer (syn. Sinorhizobium) Are Separated into Two Clades Based on Comparative Genomics and High-Throughput Phenotyping.Genome biology and evolution · 2020Article
- Use of Plasmid pVMG to Make Transcriptional ß-Glucuronidase Reporter Gene Fusions in theBiological procedures online · 2019Article
- Seven-transmembrane receptor protein RgsP and cell wall-binding protein RgsM promote unipolar growth in Rhizobiales.PLoS genetics · 2018Article
- Inter-replicon Gene Flow Contributes to Transcriptional Integration in theG3 (Bethesda, Md.) · 2018Article
- Article
- PhoU Allows Rapid Adaptation to High Phosphate Concentrations by Modulating PstSCAB Transport Rate in Sinorhizobium meliloti.Journal of bacteriology · 2017Article
- The Divided Bacterial Genome: Structure, Function, and Evolution.Microbiology and molecular biology reviews : MMBR · 2017Review
- Functional metagenomics reveals novel β-galactosidases not predictable from gene sequences.PloS one · 2017Article
- Loss of malic enzymes leads to metabolic imbalance and altered levels of trehalose and putrescine in the bacterium Sinorhizobium meliloti.BMC microbiology · 2016Article
- L-Hydroxyproline and d-Proline Catabolism in Sinorhizobium meliloti.Journal of bacteriology · 2016Article
- Cyclic Di-GMP Regulates Multiple Cellular Functions in the Symbiotic Alphaproteobacterium Sinorhizobium meliloti.Journal of bacteriology · 2016Article
- Genetic redundancy is prevalent within the 6.7 Mb Sinorhizobium meliloti genome.Molecular genetics and genomics : MGG · 2015Article
- The TyrR transcription factor regulates the divergent akr-ipdC operons of Enterobacter cloacae UW5.PloS one · 2015Article
- Genome-wide identification of CAMTA gene family members in Medicago truncatula and their expression during root nodule symbiosis and hormone treatments.Frontiers in plant science · 2015Article
- Examination of prokaryotic multipartite genome evolution through experimental genome reduction.PLoS genetics · 2014Article
Corrections and comments
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Authors and funding
9 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
As a means of investigating gene function, we developed a robust transcription fusion reporter vector to measure gene expression in bacteria. The vector, pTH1522, was used to construct a random insert library for the Sinorhizobium meliloti genome. pTH1522 replicates in Escherichia coli and can be transferred to, but cannot replicate in, S. meliloti. Homologous recombination of the DNA fragments cloned in pTH1522 into the S. meliloti genome generates transcriptional fusions to either the reporter genes gfp(+) and lacZ or gusA and rfp, depending on the orientation of the cloned fragment. Over 12,000 fusion junctions in 6,298 clones were identified by DNA sequence analysis, and the plasmid clones were recombined into S. meliloti. Reporter enzyme activities following growth of these recombinants in complex medium (LBmc) and in minimal medium with glucose or succinate as the sole carbon source allowed the identification of genes highly expressed under one or more growth condition and those expressed at very low to background levels. In addition to generating reporter gene fusions, the vector allows Flp recombinase-directed deletion formation and gene disruption, depending on the nature of the cloned fragment. We report the identification of genes essential for growth on complex medium as deduced from an inability to recover recombinants from pTH1522 clones that carried fragments internal to gene or operon transcripts. A database containing all the gene expression activities together with a web interface showing the precise locations of reporter fusion junctions has been constructed (www.sinorhizobium.org).
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