Broad host range vectors for stable genomic library construction.
We describe the construction of 36 stable vectors for genomic library construction in gram-negative species. These vectors contain the pBBR1 replicon that has been shown to stably replicate in every gram-negative species tested. The plasmids also contain bidirectional, rho-independent transcriptional terminators flanking the multiple cloning site, which allows for greater insert stability, and thus, greater genomic representation. Each vector varies in its antibiotic resistance cassette, mobilization function, and promoter used to express insert sequences. These vectors should prove useful in the screening of highly representational genomic libraries in a broad variety of gram-negative species.
Lynch
Michael D
MD
Department of Chemical and Biological Engineering, UCB424/ECCH120, University of Colorado, Boulder, 80309, USA.
Gill
Ryan T
RT
D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D024881:Drug Resistance, Bacterial / Q000235:genetics; D005798:Genes, Bacterial; D005822:Genetic Vectors; D015698:Genomic Library; D006090:Gram-Negative Bacteria / Q000235:genetics* / Q000254:growth & development; D016254:Mutagenesis, Insertional; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012093:Replicon; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA; D014158:Transcription, Genetic
D004269:DNA, Bacterial