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Updated: Aug 15, 2026

Gene-targeted Random Mutagenesis to Select Heterochromatin-destabilizing Proteasome Mutants in Fission Yeast
Published on: May 15, 2018
When coupled to natural transformation in Acinetobacter sp. strain ADP1, PCR mutagenesis is made less random by
Alison Buchan1, L Nicholas Ornston
1Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT 06520, USA.
Abstract:
Random PCR mutagenesis is a powerful tool for structure-function analysis of targeted proteins, especially when coupled with DNA integration through natural transformation followed by selection for loss of function. The technique has been applied successfully to structure-function analysis of transcriptional regulators, enzymes, and transporters in Acinetobacter sp. strain ADP1. However, the mismatch repair system prevents the full spectrum of nucleotide substitutions that may be selected at the level of protein function from being recovered. This barrier may be overcome by introducing PCR-mutagenized genes into strains in which the corresponding genes have been deleted.
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