Related Experiment Videos

Construction of a series of pACYC-derived plasmid vectors

Y Nakano1, Y Yoshida, Y Yamashita

  • 1Department of Preventive Dentistry, Kyushu University Faculty of Dentistry, Fukuoka, Japan.

Gene
|August 30, 1995
PubMed

Insights

New cloning vectors derived from pACYC177 and pACYC184 offer solutions for unstable or toxic genes. These vectors provide ampicillin or chloramphenicol resistance and are ideal for challenging gene cloning applications.

Area of Science:

  • Molecular Biology
  • Recombinant DNA Technology

Background:

  • High-copy-number vectors can lead to instability or toxicity issues when cloning certain genes.
  • Existing cloning vectors may not be suitable for handling genes with specific expression requirements.

Purpose of the Study:

  • To construct and characterize a series of novel cloning vectors.
  • To provide researchers with tools for cloning unstable and/or toxic genes.

Main Methods:

  • Construction of cloning vectors from pACYC177 and pACYC184.
  • Incorporation of a multiple cloning site (MCS), lacZ alpha reporter gene, and P15A origin of replication (ori).
  • Introduction of antibiotic resistance genes (bla for ampicillin resistance and cat for chloramphenicol resistance).

Main Results:

  • Development of plasmids pMBL18, pMBL19, pMCL200, and pMCL210 with distinct resistance markers.
  • Construction of the cosmid pMBLcos featuring the P15A ori.
  • Demonstration of vector utility for cloning genes that are unstable or toxic in high-copy-number systems.

Conclusions:

  • The newly constructed vectors are effective for cloning unstable and/or toxic genes.
  • These vectors offer versatile options for gene cloning and subcloning in molecular biology research.
  • The P15A ori and reporter gene facilitate efficient cloning strategies.

Related Concept Videos