Construction of two pGEM-7Zf(+) phagemid T-tail vectors using AhdI-restriction endonuclease sites for direct cloning of PCR products

Ji Ung Jeung, Sung K. Cho, Kyu Suk Shim, Sung Han Ok, Dae Sik Lim, Jeong Sheop Shin

Research output: Contribution to journalArticle

15 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)160-163
Number of pages4
JournalPlasmid
Volume48
Issue number2
DOIs
Publication statusPublished - 2002 Nov 20

Fingerprint

DNA Restriction Enzymes
Organism Cloning
Polymerase Chain Reaction
Genetic Vectors
beta-Lactamases
beta-Galactosidase
Transfection
Ligation
Plasmids
Color
Binding Sites
Peptides
Genes

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Construction of two pGEM-7Zf(+) phagemid T-tail vectors using AhdI-restriction endonuclease sites for direct cloning of PCR products. / Jeung, Ji Ung; Cho, Sung K.; Shim, Kyu Suk; Ok, Sung Han; Lim, Dae Sik; Shin, Jeong Sheop.

In: Plasmid, Vol. 48, No. 2, 20.11.2002, p. 160-163.

Research output: Contribution to journalArticle

Jeung, Ji Ung ; Cho, Sung K. ; Shim, Kyu Suk ; Ok, Sung Han ; Lim, Dae Sik ; Shin, Jeong Sheop. / Construction of two pGEM-7Zf(+) phagemid T-tail vectors using AhdI-restriction endonuclease sites for direct cloning of PCR products. In: Plasmid. 2002 ; Vol. 48, No. 2. pp. 160-163.
@article{8545547352e147a884ed0efc8ec1083c,
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.",
author = "Jeung, {Ji Ung} and Cho, {Sung K.} and Shim, {Kyu Suk} and Ok, {Sung Han} and Lim, {Dae Sik} and Shin, {Jeong Sheop}",
year = "2002",
month = "11",
day = "20",
doi = "10.1016/S0147-619X(02)00122-1",
language = "English",
volume = "48",
pages = "160--163",
journal = "Plasmid",
issn = "0147-619X",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Construction of two pGEM-7Zf(+) phagemid T-tail vectors using AhdI-restriction endonuclease sites for direct cloning of PCR products

AU - Jeung, Ji Ung

AU - Cho, Sung K.

AU - Shim, Kyu Suk

AU - Ok, Sung Han

AU - Lim, Dae Sik

AU - Shin, Jeong Sheop

PY - 2002/11/20

Y1 - 2002/11/20

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0036425804&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036425804&partnerID=8YFLogxK

U2 - 10.1016/S0147-619X(02)00122-1

DO - 10.1016/S0147-619X(02)00122-1

M3 - Article

VL - 48

SP - 160

EP - 163

JO - Plasmid

JF - Plasmid

SN - 0147-619X

IS - 2

ER -