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Related Experiment Videos

An E. coli-yeast shuttle cosmid with positive selection for inserted fragments.

M E Gent1, P Crowley, J R Ludwig

  • 1Department of Biochemistry and Applied Molecular Biology, University of Manchester, UK.

Current Genetics
|January 1, 1985
PubMed
Summary

A new cosmid cloning vector, pMT555, enables positive selection for inserted DNA fragments. This vector facilitates efficient genomic library construction in both E. coli and yeast, ensuring stable maintenance and replication.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Cosmid cloning vectors are essential tools for constructing genomic libraries.
  • Existing vectors may lack efficient selection mechanisms for inserted DNA.
  • Dual-host replication (E. coli and yeast) offers flexibility in molecular cloning.

Purpose of the Study:

  • To describe the development of a novel cosmid cloning vector, pMT555.
  • To enable positive selection for cloned DNA fragments.
  • To facilitate efficient genomic library construction in prokaryotic and eukaryotic systems.

Main Methods:

  • Construction of the pMT555 cosmid vector.
  • Demonstration of positive selection for inserted DNA.
  • Testing vector utility for total genomic DNA library construction.

Related Experiment Videos

  • Evaluation of replication and stable maintenance in E. coli and Saccharomyces cerevisiae.

    • Main Results:

    • The pMT555 vector allows for positive selection, confirming the presence of inserted DNA.
    • pMT555 supports replication and selection in both E. coli and S. cerevisiae.
    • Efficient construction of total genomic banks from small DNA quantities was achieved.
    • Stable maintenance of the cosmid in E. coli and faithful replication of inserts were confirmed.

    Conclusions:

    • The pMT555 cosmid vector is a valuable tool for molecular cloning and genomic library construction.
    • Positive selection enhances the reliability and efficiency of cloning experiments.
    • The dual-host capability broadens the applicability of pMT555 in genetic research.