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Isolation and use of Prevotella ruminicola TC18 plasmid pTC18 in Escherichia coli-P. ruminicola shuttle vector construction
Accetto, Tomaž (Author)

URLURL - Presentation file, Visit http://aas.bf.uni-lj.si/zootehnika/94-2009/PDF/94-2009-2-143-146.pdf This link opens in a new window

Abstract
A cryptic plasmid of approximately 3 kilobases named pTC18 was discovered in a ruminal Prevotella ruminicola TC18 strain and cloned into Escherichia coli. Based on pTC18, several shuttle vectors, containing Prevotella/Bacteroides tetQ selection marker and E. coli vector pUC19 inserted at two different positions in pTC18 were constructed. The shuttle vectors, protected with HaeIII methylase against the P. ruminicola 23 restriction were electroporated into P. ruminicola. Despite numerous attempts a tetracycline resistant recombinant strain 23 was not obtained. The possible causes for electroporation failure are discussed.

Language:English
Keywords:microbiology, anaerobic bacteria, Prevotella ruminicola, shuttler vector, rumen
Work type:Not categorized (r6)
Tipology:1.01 - Original Scientific Article
Organization:BF - Biotechnical Faculty
Year:2009
Number of pages:str. 143-146
Numbering:Letn.94, št. 2
UDC:579
ISSN on article:1581-9175
COBISS.SI-ID:2546824 Link is opened in a new window
Views:643
Downloads:207
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Record is a part of a journal

Title:Acta agriculturae Slovenica
Shortened title:Acta agric. Slov.
Publisher:Biotehniška fakulteta
ISSN:1581-9175
COBISS.SI-ID:213840640 This link opens in a new window

Secondary language

Language:Slovenian
Abstract:
V vampnem sevu Prevotella ruminicola TC18 smo odkrili 3 kilobazne pare dolgo plazmidno DNA, jo poimenovali pTC18 in klonirali v Escherichia coli. Na njeni osnovi smo razvili več različic prenosljivega plazmida, ki je poleg pTC18 vseboval še selekcijski marker tetQ iz sevov rodu Bacteroides in plazmidni vektor E. coli pUC19. Prenosljive vektorje smo s HaeIII metilazo zaščitili proti restrikciji v P. ruminicola 23 in jih nato poskusili vnesti v P. ruminicola 23 z elektrotransformacijo. Kljub mnogim poskusom nismo uspeli pridobiti proti tetraciklinu odpornih sevov P. ruminicola 23.

Keywords:mikrobiologija, anaerobne bakterije, Prevotella rumiciola, prenosljivi vektor, vamp

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