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Related Experiment Videos

Generalized and mobilizable positive-selection cloning vectors

S Mongkolsuk1, S Rabibhadana, P Vattanaviboon

  • 1Laboratory of Biotechnology, Chulabhorn Research Institute, Lak si, Bangkok, Thailand.

Gene
|May 27, 1994
PubMed
Summary

Researchers enhanced the pUN121 cloning vector, creating pSKM10 with more unique cloning sites. A mobilizable version, pSKM11, allows easy transfer into various Gram-negative bacteria.

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Area of Science:

  • Molecular Biology
  • Bacteriology

Background:

  • The pUN121 plasmid is a positive-selection cloning vector.
  • Enhancing cloning vectors with more unique restriction sites is crucial for molecular biology applications.

Purpose of the Study:

  • To modify the pUN121 cloning vector to increase the number of unique cloning sites.
  • To create a mobilizable version of the modified vector for broad bacterial host applications.

Main Methods:

  • Insertion of a multiple cloning site (MCS) into the lambda cI gene of pUN121 without disrupting its repressor function, creating pSKM10.
  • Addition of the IncP mob sequence to pSKM10 to generate the mobilizable plasmid pSKM11.

Main Results:

  • Plasmid pSKM10 possesses seven distinct restriction enzyme sites for general cloning.

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  • Plasmid pSKM11 demonstrated successful mobilization into a wide range of Gram-negative bacteria.

    • Conclusions:

    • The modified pSKM10 vector offers expanded cloning capabilities.
    • The mobilizable pSKM11 plasmid facilitates efficient genetic transfer across diverse Gram-negative bacterial species.