title: Construction of two pGEM-7Zf(+) phagemid T-tail vectors using AhdI-restriction endonuclease sites for direct cloning of PCR products.
abstract: For applications such as sequencing, transfection, and in vitro transcription, PCR products have to be subcloned into plasmids. Many strategies are used for cloning, blunt-end ligation or the incorporation of restriction endonuclease sites into PCR primers for appropriate vectors. However, the most convenient and direct method is T/A cloning. In this study, we developed two of the pGEM-7Zf(+) phagemid T-tail vectors using AhdI-restriction endonuclease sites, and these T vectors have all the features of pGEM-7Zf(+): f1 ori, T7, and SP6 RNA polymerase promoters, the alpha-peptide coding region of beta-galactosidase for X-gal blue/white color selection, the beta-lactamase gene for recombinant colony selection, and binding sites for pUC/M13 forward and reverse sequencing primers. These AhdI-containing phagemid vectors, pGEM-NJ105 and pGEM-NJ107, are useful for the easy and inexpensive preparation of T vectors and direct cloning of PCR products.
authors.affiliation: Graduate School of Biotechnology, Korea University, Seoul 136-701, South Korea.
authors.forename: Ji Ung
authors.initials: JU
authors.lastname: Jeung
authors.forename: Sung Ki
authors.initials: SK
authors.lastname: Cho
authors.forename: Kyu Suk
authors.initials: KS
authors.lastname: Shim
authors.forename: Sung Han
authors.initials: SH
authors.lastname: Ok
authors.forename: Dae Sik
authors.initials: DS
authors.lastname: Lim
authors.forename: Jeong Sheop
authors.initials: JS
authors.lastname: Shin
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D005822:Genetic Vectors / Q000235:genetics*; D016133:Polymerase Chain Reaction; D013379:Substrate Specificity; D014170:Transformation, Genetic
chemical_list: D004262:DNA Restriction Enzymes