Abstract
The dissimilatory sulfate-reducing deltaproteobacterium Desulfovibrio vulgaris Hildenborough (ATCC 29579) was chosen by the research collaboration ENIGMA to explore tools and protocols for bringing this anaerobe to model status. Here, we describe a collection of genetic constructs generated by ENIGMA that are available to the research community.
Highlights
Because of the environmental importance of these microbes, the first sulfate-reducing bacterium with a sequenced genome, Desulfovibrio vulgaris Hildenborough [7], was chosen to be brought to model status for use in generating a transposon (Tn) library and for constructing strains that produce affinity-tagged proteins (Table 1)
Marker exchange mutation, replacing a nucleotide sequence with a selectable marker flanked by homologous chromosomal regions, has been the cornerstone of genetic constructions [10, 11]
To generate in-frame deletions without a residual selectable marker, a parental strain, JW710, was created that is resistant to inhibition by 5fluorouracil through deletion of the uracil phosphoribosyltransferase gene [12]
Summary
Because of the environmental importance of these microbes, the first sulfate-reducing bacterium with a sequenced genome, Desulfovibrio vulgaris Hildenborough [7], was chosen to be brought to model status for use in generating a transposon (Tn) library and for constructing strains that produce affinity-tagged proteins (Table 1). The plasmids, pSC27 [15], pMO719 [16], pMO9075 [11], and pMO746 [17], with features used in various strain constructions, are available from https://www.addgene.org/Judy_Wall/. Citation Wall JD, Zane GM, Juba TR, Kuehl JV, Ray J, Chhabra SR, Trotter VV, Shatsky M, De León KB, Keller KL, Bender KS, Butland G, Arkin AP, Deutschbauer AM.
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