Extremophiles

, Volume 9, Issue 6, pp 427–435

Gene replacement in Haloarcula marismortui: construction of a strain with two of its three chromosomal rRNA operons deleted

Authors

  • Daqi Tu
    • Department of Molecular Biophysics and BiochemistryYale University
  • Gregor Blaha
    • Department of Molecular Biophysics and BiochemistryYale University
    • Department of Molecular Biophysics and BiochemistryYale University
    • Department of ChemistryYale University
  • Thomas A. Steitz
    • Department of Molecular Biophysics and BiochemistryYale University
    • Department of ChemistryYale University
    • Howard Hughes Medical Institute
Original Paper

DOI: 10.1007/s00792-005-0459-y

Cite this article as:
Tu, D., Blaha, G., Moore, P.B. et al. Extremophiles (2005) 9: 427. doi:10.1007/s00792-005-0459-y

Abstract

Site-directed mutagenesis were done in Haloarcula marismortui using the strategy that Khorana and coworkers devised for deleting the bacteriorhodopsin gene from Halobacterium halobium [Krebs et al. Proc Natl Acad Sci USA 90:1987–1991 (1993)]. Strains have been prepared from H. marsimortui, which normally has three rRNA operons, that are missing either its rrnB operon or both its rrnB and rrnC operons. In rich media, both strains grow at about the same rate as wild type. The G2099 in the 23S rRNA gene of the single operon strain was changed to A, and a three amino acid deletion was introduced into the gene for ribosomal protein L22 of the wild-type organism. The structural consequences of these and other such mutations can be determined with unusual accuracy because crystals of the large ribosomal subunit of H. marismortui diffract to atomic resolution.

Keywords

Haloarcula marismortuiGene replacementrRNA OperonRibosomal protein L22

Abbreviations

rrnA

ribosomal RNA operon A

rrnB

ribosomal RNA operon B

rrnC

ribosomal RNA operon C

rRNA

ribosomal RNA

Mevr

Mevinolin resistant

Copyright information

© Springer-Verlag 2005