λ Recombineering Used to Engineer the Genome of Phage T7

Antibiotics
Jordan D JensenDonald L Court

Abstract

Bacteriophage T7 and T7-like bacteriophages are valuable genetic models for lytic phage biology that have heretofore been intractable with in vivo genetic engineering methods. This manuscript describes that the presence of λ Red recombination proteins makes in vivo recombineering of T7 possible, so that single base changes and whole gene replacements on the T7 genome can be made. Red recombination functions also increase the efficiency of T7 genome DNA transfection of cells by ~100-fold. Likewise, Red function enables two other T7-like bacteriophages that do not normally propagate in E. coli to be recovered following genome transfection. These results constitute major technical advances in the speed and efficiency of bacteriophage T7 engineering and will aid in the rapid development of new phage variants for a variety of applications.

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Citations

Apr 22, 2021·BioDrugs : Clinical Immunotherapeutics, Biopharmaceuticals and Gene Therapy·Małgorzata ŁobockaAndrzej Górski
Jul 30, 2021·Biochimica Et Biophysica Acta. General Subjects·Monika ŠimoliūnienėLidija Truncaitė

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Methods Mentioned

BETA
Transfection
PCR
genetic modification

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