A Modular, Positive Selection Bacterial Artificial Chromosome Vector with Multiple Cloning Sites

ABSTRACT Bacterial artificial chromosome (BAC) vectors are important tools for microbial genome research. We constructed a novel BAC vector, pUvBBAC, for replication in both gram-negative and gram-positive bacterial hosts. The pUvBBAC vector was used to generate a BAC library for the facultative intracellular pathogen Listeria monocytogenes EGD-e. The library had insert sizes ranging from 68 to 178 kb. We identified two recombinant BACs from the L. monocytogenes pUvBBAC library that each contained the entire virulence gene cluster (vgc) of L. monocytogenes and transferred them to a nonpathogenic Listeria innocua strain. Recombinant L. innocua strains harboring pUvBBAC+vgc1 and pUvBBAC+vgc2 produced the vgc-specific listeriolysin (LLO) and actin assembly protein ActA and represent the first reported cloning of the vgc locus in its entirety. The use of the novel broad-host-range BAC vector pUvBBAC extends the versatility of this technology and provides a powerful platform for detailed functional genomics of gram-positive bacteria as well as its use in explorative functional metagenomics.

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Small-Scale Preparation of the Single-Copy Bacterial Artificial Chromosome Vector pBeloBAC11

BioTechniques ◽

10.2144/98244bm10 ◽

1998 ◽

Vol 24 (4) ◽

pp. 568-572 ◽

Cited By ~ 6

Author(s):

Daniel Tillett ◽

Brett A. Neilan

Keyword(s):

Bacterial Artificial Chromosome ◽

Artificial Chromosome ◽

Single Copy ◽

Small Scale ◽

Bacterial Artificial Chromosome Vector ◽

Scale Preparation

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Faculty Opinions recommendation of Cloning the vaccinia virus genome as a bacterial artificial chromosome in Escherichia coli and recovery of infectious virus in mammalian cells.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1009413.126057 ◽

2002 ◽

Author(s):

Grant McFadden

Keyword(s):

Escherichia Coli ◽

Bacterial Artificial Chromosome ◽

Vaccinia Virus ◽

Mammalian Cells ◽

Infectious Virus ◽

Artificial Chromosome ◽

Virus Genome

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Targeting CreERT2 expression to keratin 8-expressing murine simple epithelia using bacterial artificial chromosome transgenesis

Transgenic Research ◽

10.1007/s11248-012-9598-y ◽

2012 ◽

Vol 21 (5) ◽

pp. 1117-1123 ◽

Cited By ~ 15

Author(s):

Li Zhang ◽

Boyu Zhang ◽

Sang Jun Han ◽

Amy N. Shore ◽

Jeffrey M. Rosen ◽

...

Keyword(s):

Bacterial Artificial Chromosome ◽

Artificial Chromosome ◽

Keratin 8

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Revising the Chromosome-Specific Probes of White Hawk (Leucopternis albicollis)

Proceedings ◽

10.3390/iecge-07152 ◽

2020 ◽

Vol 76 (1) ◽

pp. 1

Author(s):

Ivanete Furo ◽

Rafael Kretschmer ◽

Jorge Pereira ◽

Darren Griffin ◽

Rebecca O’Connor ◽

...

Keyword(s):

Bacterial Artificial Chromosome ◽

Artificial Chromosome ◽

Bird Of Prey ◽

Inaccurate Data ◽

Intrachromosomal Rearrangements

Leucopternis albicollis is a diurnal bird of prey with extensive karyotype reorganization. Chromosome-specific probes from this species have been used successfully to detect intrachromosomal rearrangements in different species of bird since 2010. However, some gaps were detected in this first set of probes. Here, we have obtained a new set of whole chromosome probes in order to improve the previous one; also, we have performed experiments using bacterial artificial chromosome (BAC) from chicken microchromosomes. Our results demonstrated that the microchromosomes were involved in fusion events. In addition, a new nomenclature has been proposed for the new set of probes and some inaccurate data were corrected.

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