title: A modular, positive selection bacterial artificial chromosome vector with multiple cloning sites.
abstract: To construct large-insert libraries for the sequencing, mapping, and functional studies of complex genomes, we have constructed a new modular bacterial artificial chromosome (BAC) vector, pBACe3.6 (GenBank Accession No. U80929). This vector contains multiple cloning sites located within the sacB gene, allowing positive selection for recombinant clones on sucrose-containing medium. A recognition site for the PI-SceI nuclease has also been included, which permits linearization of recombinant DNA irrespective of the characteristics of the insert sequences. An attTn7 sequence present in pBACe3.6 permits retrofitting of BAC clones by Tn7-mediated insertion of desirable sequence elements into the vector portion. The ability to retrofit BAC clones will be useful for functional analysis of genes carried on the cloned inserts. The pBACe3.6 vector has been used for the construction of many genomic libraries currently serving as resources for large-scale mapping and sequencing.
authors.affiliation: Department of Cancer Genetics, Roswell Park Cancer Institute, Buffalo, New York 14263, USA.
authors.forename: E
authors.initials: E
authors.lastname: Frengen
authors.forename: D
authors.initials: D
authors.lastname: Weichenhan
authors.forename: B
authors.initials: B
authors.lastname: Zhao
authors.forename: K
authors.initials: K
authors.lastname: Osoegawa
authors.forename: M
authors.initials: M
authors.lastname: van Geel
authors.forename: P J
authors.initials: PJ
authors.lastname: de Jong
mesh_terms: D001665:Binding Sites; D002876:Chromosomes, Bacterial / Q000235:genetics*; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D008969:Molecular Sequence Data
chemical_list: D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D004262:DNA Restriction Enzymes