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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1
pubmed:dateCreated
1994-12-21
pubmed:abstractText
A detailed examination of vectors and procedures used for Tn5 mutagenesis of the phototrophic purple non-sulfur bacterium Rhodospirillum rubrum has been performed. The mobilizable Tn5 suicide vectors currently available show a frequency of Tn5 mutagenesis for R. rubrum of approx. 10(-7)-10(-8), approx. 100-1000-fold lower than observed for the related bacteria Rhodobacter capsulatus and Rhodobacter sphaeroides. Using the blue-to-red reversion of a blue-green mutant, R. rubrum ST6, containing a single Tn5 lesion in one of the early genes for carotenoid biosynthesis, we have shown that the frequency of precise excision of a chromosomally inserted Tn5 element, to restore the wild-type phenotype in the absence of selection, is 10(-6). We have constructed three new suicide vectors for Tn5 mutagenesis, where the transposase encoded by the IS50R element was placed in the same (pSUPEG11, pSUPEG21) or in the opposing (pSUPEG22) orientation from the weak promoter of the RK2-derived tetR gene. With the vector pSUPEG11, the frequency of Tn5 mutagenesis was increased to 10(-5), approx. 100-fold higher than observed previously.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
0378-1119
pubmed:author
pubmed:issnType
Print
pubmed:day
2
pubmed:volume
150
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
97-100
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed:year
1994
pubmed:articleTitle
An improved procedure and new vectors for transposon Tn5 mutagenesis of the phototrophic bacterium Rhodospirillum rubrum.
pubmed:affiliation
Department of Microbiology, Biozentrum, Basel, Switzerland.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't