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. IVR 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 AhaI-restriction endonuclease sites. and these T vectors have all the features of pGFM-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-NJI07. are useful Cot, the easy and inexpensive preparation of T vectors and direct cloning of PCR products. (C) 2002 Elsevier Science (USA). All rights reserved.