|Title||Two-Step Bacterial Artificial Chromosome (BAC) Engineering: Preparation of Shuttle Vector DNA.|
|Publication Type||Journal Article|
|Year of Publication||2020|
|Authors||Heintz N, Gong S|
|Journal||Cold Spring Harb Protoc|
|Date Published||2020 04 01|
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.
|Alternate Journal||Cold Spring Harb Protoc|