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Related Experiment Videos

Bacteriophage lambda as a cloning vector.

V M Chauthaiwale1, A Therwath, V V Deshpande

  • 1Division of Biochemical Sciences, National Chemical Laboratory, Pune, India.

Microbiological Reviews
|December 1, 1992
PubMed
Summary

Lambda vectors have been extensively modified for gene cloning since the 1970s. These versatile tools facilitate DNA manipulation, cloning, and analysis, making them popular in molecular biology research.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Bacteriophage lambda has been a key tool in molecular cloning since the late 1970s.
  • Understanding lambda's molecular organization and genetic regulation enabled its modification for gene manipulation.

Purpose of the Study:

  • To review the development and applications of multipurpose lambda vectors for molecular cloning.
  • To highlight the diverse modifications and features that enhance lambda vectors' utility.

Main Methods:

  • Modification of lambda genome to alter restriction sites and incorporate polylinkers.
  • Development of in vitro packaging systems for efficient DNA introduction.
  • Engineering vectors with specific features like amber mutations for biological containment.
  • Design of vectors accommodating various insert sizes (up to 24 kbp).

Main Results:

  • Creation of diverse lambda vectors (e.g., Charon, EMBL, lambda gt10, lambda gt11, lambda gt22, lambda DASH, lambda ZAP) with tailored properties.
  • Vectors facilitate cloning of different DNA fragment sizes and offer features for in vitro transcription and in vivo excision.
  • Recombinants are easily distinguishable from parental phages.
  • Lambda vector libraries allow efficient screening using various probes.

Conclusions:

  • Lambda vectors, enhanced by efficient packaging and containment strategies, are powerful tools for molecular cloning.
  • Adaptations such as specific polylinkers, promoters, and excision mechanisms have significantly improved cloning and analysis.
  • The combination of vector availability and supporting techniques solidifies lambda's role in modern molecular biology.

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