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Two-Step Bacterial Artificial Chromosome (BAC) Engineering: Preparation of Shuttle Vector DNA.

TitleTwo-Step Bacterial Artificial Chromosome (BAC) Engineering: Preparation of Shuttle Vector DNA.
Publication TypeJournal Article
Year of Publication2020
AuthorsHeintz N, Gong S
JournalCold Spring Harb Protoc
Volume2020
Issue4
Pagination098038
Date Published2020 04 01
ISSN1559-6095
Abstract

In two-step bacterial artificial chromosome (BAC) engineering, a single plasmid is introduced into the BAC-carrying cell lines. The shuttle vector pLD53.SCAB (or pLD53.SCAEB) carries the gene and the R6Kγ origin, which requires the π protein to replicate. PIR2 cells, expressing π, are typically used for the amplification of the vector and maintain about 15 copies/cell of the donor vector, which is relatively stable in this host.

DOI10.1101/pdb.prot098038
Alternate JournalCold Spring Harb Protoc
PubMed ID32238589