Establishing a Markerless Genetic Exchange System for Methanosarcina mazei Strain Gö1 for Constructing Chromosomal Mutants of Small RNA Genes
Joint Authors
Ehlers, Claudia
Schmitz, Ruth A.
Jäger, Dominik
Source
Issue
Vol. 2011, Issue 2011 (31 Dec. 2011), pp.1-7, 7 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2011-09-20
Country of Publication
Egypt
No. of Pages
7
Main Subjects
Abstract EN
A markerless genetic exchange system was successfully established in Methanosarcina mazei strain Gö1 using the hpt gene coding for hypoxanthine phosphoribosyltransferase.
First, a chromosomal deletion mutant of the hpt gene was generated conferring resistance to the purine analog 8-aza-2,6-diaminopurine (8-ADP).
The nonreplicating allelic exchange vector (pRS345) carrying the pac-resistance cassette for direct selection of chromosomal integration, and the hpt gene for counterselection was introduced into this strain.
By a pop-in and ultimately pop-out event of the plasmid from the chromosome, allelic exchange is enabled.
Using this system, we successfully generated a M.
mazei deletion mutant of the gene encoding the regulatory non-coding RNA sRNA154.
Characterizing M.
mazeiΔsRNA154 under nitrogen limiting conditions demonstrated differential expression of at least three cytoplasmic proteins and reduced growth strongly arguing for a prominent role of sRNA154 in regulation of nitrogen fixation by posttranscriptional regulation.
American Psychological Association (APA)
Ehlers, Claudia& Jäger, Dominik& Schmitz, Ruth A.. 2011. Establishing a Markerless Genetic Exchange System for Methanosarcina mazei Strain Gö1 for Constructing Chromosomal Mutants of Small RNA Genes. Archaea،Vol. 2011, no. 2011, pp.1-7.
https://search.emarefa.net/detail/BIM-472469
Modern Language Association (MLA)
Ehlers, Claudia…[et al.]. Establishing a Markerless Genetic Exchange System for Methanosarcina mazei Strain Gö1 for Constructing Chromosomal Mutants of Small RNA Genes. Archaea No. 2011 (2011), pp.1-7.
https://search.emarefa.net/detail/BIM-472469
American Medical Association (AMA)
Ehlers, Claudia& Jäger, Dominik& Schmitz, Ruth A.. Establishing a Markerless Genetic Exchange System for Methanosarcina mazei Strain Gö1 for Constructing Chromosomal Mutants of Small RNA Genes. Archaea. 2011. Vol. 2011, no. 2011, pp.1-7.
https://search.emarefa.net/detail/BIM-472469
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-472469