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PMID: 11844777 Published · ppublish English Evaluation Study Journal Article Research Support, Non-U.S. Gov't

Directed mutagenesis and plasmid-based complementation in the methanogenic archaeon Methanosarcina acetivorans C2A demonstrated by genetic analysis of proline biosynthesis.

Journal of bacteriology ·Vol. 184 ·No. 5 ·2002-03-00 ·Pages 1449-54

Zhang JK, White AK, Kuettner HC, Boccazzi P, Metcalf WW

Abstract

We report here the first use of directed mutagenesis in Methanosarcina acetivorans C2A. The method employs homologous recombination-mediated gene replacement and was used to construct a variety of proline auxotrophs with mutations in the proABC locus. Each mutation was also complemented in trans with autonomously replicating Methanosarcina-Escherichia plasmid shuttle vectors.

MeSH Terms
Aldehyde Oxidoreductases/genetics Archaeal Proteins/genetics,metabolism Cloning, Molecular Genes, Archaeal Genetic Complementation Test/methods Glutamate-5-Semialdehyde Dehydrogenase Methanosarcina/genetics,metabolism Molecular Sequence Data Mutagenesis, Insertional/methods Phosphotransferases (Carboxyl Group Acceptor)/genetics Plasmids/genetics Proline/biosynthesis Pyrroline Carboxylate Reductases/genetics Recombination, Genetic Sequence Analysis, DNA
Chemicals
Archaeal Proteins Proline Aldehyde Oxidoreductases Glutamate-5-Semialdehyde Dehydrogenase Pyrroline Carboxylate Reductases Phosphotransferases (Carboxyl Group Acceptor) glutamate 5-kinase
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Zhang Jun Kai
Department of Microbiology, B103 Chemical and Life Science Laboratory, University of Illinois, 601 S. Goodwin Ave., Urbana, IL 61801, USA.
White Andrea K
Kuettner H Craig
Boccazzi Paolo
Metcalf William W
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/JB.184.5.1449-1454.2002
Published
2002-03-00
Pages
1449-54
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC134853
Subset
IM
Databases
GENBANK
AF305580
External Links
PubMed source DOI: 10.1128/JB.184.5.1449-1454.2002 PMC Full Text View journal details
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