PMID: 2121708Nov 1, 1990Paper

Gene-scrambling mutagenesis: generation and analysis of insertional mutations in the alginate regulatory region of Pseudomonas aeruginosa

Journal of Bacteriology
C D Mohr, V Deretic

Abstract

A novel method for random mutagenesis of targeted chromosomal regions in Pseudomona aeruginosa was developed. This method can be used with a cloned DNA fragment of indefinite size that contains a putative gene of interest. Cloned DNA is digested to produce small fragments that are then randomly reassembled into long DNA inserts by using cosmid vectors and lambda packaging reaction. This DNA is then transferred into P. aeruginosa and forced into the chromosome via homologous recombination, producing in a single step a random set of insertional mutants along a desired region of the chromosome. Application of this method to extend the analysis of the alginate regulatory region, using a cloned 6.2-kb fragment with the algR gene and the previously uncharacterized flanking regions, produced several insertional mutations. One mutation was obtained in algR, a known transcriptional regulatory of mucoidy in P. aeruginosa. The null mutation of algR was generated in a mucoid derivative of the standard genetic strain PAO responsive to different environmental factors. This mutation was used to demonstrate that the algR gene product was not essential for the regulation of its promoters. Additional insertions were obtained in regions downstrea...Continue Reading

References

Dec 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·F SangerA R Coulson
Apr 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·D H Figurski, D R Helinski
Jul 7, 1977·Molecular & General Genetics : MGG·D HaasT Leisinger
Feb 1, 1990·Journal of Bacteriology·V Stout, S Gottesman
Oct 31, 1989·Biochemical and Biophysical Research Communications·K Kimbara, A M Chakrabarty
Sep 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·B AricóR Rappuoli
Dec 1, 1989·Microbiological Reviews·J B StockA M Stock
Mar 1, 1989·Proceedings of the National Academy of Sciences of the United States of America·K S Ishimoto, S Lory
Apr 21, 1989·Cell·J Piatigorsky, G J Wistow
Oct 11, 1988·Nucleic Acids Research·S E West, B H Iglewski
Mar 22, 1985·Science·D J Lipman, W R Pearson
Feb 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·S Tabor, C C Richardson
Mar 11, 1988·Nucleic Acids Research·J M Pustell
Jun 19, 1987·Science·G Wistow, J Piatigorsky
Nov 1, 1985·Journal of Bacteriology·A DarzinsA M Chakrabarty
Dec 1, 1972·Journal of General Microbiology·J H Isaac, B W Holloway
Sep 1, 1968·Analytical Biochemistry·C A Knutson, A Jeanes
Apr 1, 1981·Analytical Biochemistry·G DretzenP Chambon
Jan 1, 1983·Methods in Enzymology·J Messing
Jul 1, 1983·Analytical Biochemistry·A P Feinberg, B Vogelstein

❮ Previous
Next ❯

Citations

Dec 10, 2014·Journal of Bacteriology·Christopher L PritchettMichael J Schurr
Sep 11, 1999·FEMS Microbiology Letters·C OlveraG Soberón-Chávez
Jul 8, 2014·Frontiers in Cellular and Infection Microbiology·Yuta OkkotsuMichael J Schurr
May 29, 1999·Journal of Bacteriology·M K FakhrC L Bender

❮ Previous
Next ❯

Related Concepts

Related Feeds

Biosynthetic Transformations

Biosyntheic transformtions are multi-step, enzyme-catalyzed processes where substrates are converted into more complex products in living organisms. Simple compounds are modified, converted into other compounds, or joined together to form macromolecules. Discover the latest research on biosynthetic transformations here.