Developing Genetic Tools for the Planctomycetes and ...

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Developing Genetic Tools for the Planctomycetes and Verrucomicrobia: Novel approaches to transformation

Name:Personal
Domman, D.
Role :Text(marcrelator)
creator

Name:Personal
Ward, N.L.
Role :Text(marcrelator)
creator

Name:Personal
Ward, B.
Role :Text(marcrelator)
creator

Name:Personal
Sait, M.
Role :Text(marcrelator)
creator

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still image
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Laramie, Wyoming

University of Wyoming
(keyDate="yes")
2009-05-13

Language:Text
eng

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born digtal

abstract
The phyla Planctomycetes and Verrucomicrobia present a unique set of characteristics that differentiate them from other prokaryotes. These include an altered cell wall structure lacking peptidoglycan, the presence of an intracytoplasmic membrane, nuclear body envelope, and sterol synthesis. Superficially, some of these characteristics resemble those in eukaryotes, raising questions of their origin and function. However, these phyla are relatively poorly characterized and lack the molecular tools needed for further characterization. The initial experiment tested the effect of different electroporation conditions and buffer solutions on uptake of foreign DNA by Verrucomicrobium spinosum and Gemmata obscuriglobus. Presented here is a novel approach to the transformation of G. obscuriglobus via electroporation using a 10% sucrose buffering solution. Preliminary evidence indicates that an antibiotic resistance encoding Tn5 transposon carried on plasmid pAG408 was integrated into the host genome. This integration permitted growth on a kanomycin-supplemented medium but did not support growth on a gentomycin-supplemented medium. To date, 37 putative mutants have been identified and are currently being screened by PCR to identify insertion sites. Future work includes the utilization of this electroporation protocol to generate targeted mutations and a stable expression system. These methodologies will hopefully increase the molecular tools available to study these unique organisms.
note
From - Undergraduate Research Day 2009 - Celebration of Research - Abstracts
Subject
Planctomycetes

Subject
Verrucomicrobia

Related Item:series Title Information
Undergrauate Research Day 2009

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http://digital.uwyo.edu/copyright.htm
Record Information languageOfCataloging :Text(ISO639-2B)
English
:Code(ISO639-2B)
eng