Exploiting a natural auxotrophy for genetic selection.
We exploited the natural histidine auxotrophy of Francisella species to develop hisD (encodes histidinol dehydrogenase) as a positive selection marker. A shuttle plasmid (pBR103) carrying Escherichia coli hisD and designed for cloning of PCR fragments replicated in both attenuated and highly virulent Francisella strains. During this work, we formulated a simplified defined growth medium for Francisella novicida.
Department of Genome Sciences, University of Washington, Seattle, Washington, USA.
Elizabeth
E
Ramage
Larry
L
Gallagher
Colin
C
Manoil
D000429:Alcohol Oxidoreductases / Q000235:genetics / Q000378:metabolism; D003470:Culture Media / Q000737:chemistry; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005603:Francisella / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development* / Q000378:metabolism; D005818:Genetic Engineering / Q000379:methods; D005822:Genetic Vectors; D005827:Genetics, Microbial / Q000379:methods*; D006639:Histidine / Q000378:metabolism; D008969:Molecular Sequence Data; D010957:Plasmids; D012641:Selection, Genetic; D017422:Sequence Analysis, DNA
D003470:Culture Media; D004269:DNA, Bacterial; D006639:Histidine; D000429:Alcohol Oxidoreductases; C021358:histidinol dehydrogenase