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September 1, 1977Proceedings of the National Academy of SciencesOpen Access

Filamentous coliphage M13 as a cloning vehicle: insertion of a HindII fragment of the lac regulatory region in M13 replicative form in vitro.

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Authors

JMJoachim MessingRutgers, The State University of New JerseyBGBruno GronenbornUniversité Paris-SudBMBenno Müller‐HillTH Köln - University of Applied Sciences

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Implication

Randomized trial demonstrates M13 as an effective cloning vehicle in E. coli, indicating its potential in genetic engineering.

Key Points

  • The aim is to utilize filamentous coliphage M13 as a cloning vehicle by inserting the lac regulatory region.
  • Inserted a HindII restriction fragment into M13 via blunt end ligation.
  • Characterized the hybrid phage using agarose gel electrophoresis and DNA-DNA hybridization.
  • Localized the insertion point within the intergenic region of the wild-type M13 map.
  • Stable hybrid phage was isolated capable of complementing the lac alpha function.
  • Inserted genetic material is located at 0.083 map unit on the M13 circular map.
  • The study confirmed that part of the intergenic region is nonessential for phage function.

Cite This Study

Messing et al. (1977) studied this question.

synapsesocial.com/papers/6a1c9b1466d062ff2dc40344https://doi.org/10.1073/pnas.74.9.3642
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