| 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 |
| Volume | 2020 |
| Issue | 4 |
| Pagination | 098038 |
| Date Published | 2020 04 01 |
| ISSN | 1559-6095 |
| 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. |
| DOI | 10.1101/pdb.prot098038 |
| Alternate Journal | Cold Spring Harb Protoc |
| PubMed ID | 32238589 |