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Plasmids for recombination-based screening.

Publication ,  Journal Article
Stewart, GD; Hauser, MA; Kang, H; McCann, DP; Osemlak, MM; Kurnit, DM; Hanzlik, AJ
Published in: Gene
September 30, 1991

To facilitate recombination-based screening, we constructed the ColE1-based plasmid, pi G4, that confers chloramphenicol resistance, contains a polylinker with multiple unique restriction enzyme recognition sequences, and contains the genetic marker, supF. To facilitate recombination-based screening followed by rapid DNA sequencing, we inserted the selectable marker, supF, into each of 20 high-copy-number (hcn) pUC-derived NoC plasmids that were designed for multiplex DNA sequencing. To facilitate recombination-based screening of common cDNA libraries that often contain ColE1 sequences, we constructed a supF-carrying plasmid whose replication was driven from an R6K replicon that does not share sequence homology with ColE1. Furthermore, we incorporated a useful polylinker and increased the copy number of this plasmid to create the 4.4-kb hcn plasmid, pMAD1. Thus, these plasmids allow: (1) background-free transformation of cells by a supF plasmid carrying an antibiotic-resistance marker; (2) simultaneous performance of the recombination-based assay and DNA sequencing; and (3) screening bacteriophage cDNA libraries that contain ColE1 sequences by recombination with a supF plasmid that is not homologous to ColE1 derivatives.

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Published In

Gene

DOI

ISSN

0378-1119

Publication Date

September 30, 1991

Volume

106

Issue

1

Start / End Page

97 / 101

Location

Netherlands

Related Subject Headings

  • Replicon
  • Recombination, Genetic
  • Plasmids
  • Oligodeoxyribonucleotides
  • Molecular Sequence Data
  • Genetic Markers
  • Genes, Viral
  • Developmental Biology
  • DNA, Viral
  • DNA
 

Citation

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Stewart, G. D., Hauser, M. A., Kang, H., McCann, D. P., Osemlak, M. M., Kurnit, D. M., & Hanzlik, A. J. (1991). Plasmids for recombination-based screening. Gene, 106(1), 97–101. https://doi.org/10.1016/0378-1119(91)90571-r
Stewart, G. D., M. A. Hauser, H. Kang, D. P. McCann, M. M. Osemlak, D. M. Kurnit, and A. J. Hanzlik. “Plasmids for recombination-based screening.Gene 106, no. 1 (September 30, 1991): 97–101. https://doi.org/10.1016/0378-1119(91)90571-r.
Stewart GD, Hauser MA, Kang H, McCann DP, Osemlak MM, Kurnit DM, et al. Plasmids for recombination-based screening. Gene. 1991 Sep 30;106(1):97–101.
Stewart, G. D., et al. “Plasmids for recombination-based screening.Gene, vol. 106, no. 1, Sept. 1991, pp. 97–101. Pubmed, doi:10.1016/0378-1119(91)90571-r.
Stewart GD, Hauser MA, Kang H, McCann DP, Osemlak MM, Kurnit DM, Hanzlik AJ. Plasmids for recombination-based screening. Gene. 1991 Sep 30;106(1):97–101.
Journal cover image

Published In

Gene

DOI

ISSN

0378-1119

Publication Date

September 30, 1991

Volume

106

Issue

1

Start / End Page

97 / 101

Location

Netherlands

Related Subject Headings

  • Replicon
  • Recombination, Genetic
  • Plasmids
  • Oligodeoxyribonucleotides
  • Molecular Sequence Data
  • Genetic Markers
  • Genes, Viral
  • Developmental Biology
  • DNA, Viral
  • DNA