Environmental performance of Pseudomonas putida with a uracylated genome.
Environmental performance of Pseudomonas putida with a uracylated genome.
Chembiochem. 2020 Jun 29;:
Authors: Algar E, Al-Ramahi Y, de Lorenzo V, Martínez-García E
Abstract
A variant of the soil bacterium Pseudomonas putida with a genome containing a ∼ 20% replacement of the whole of thymine (T) by uracil (U) was made by deleting genes ung (uracil DNA glycosylase) and dut ( deoxyuridine 5'-triphosphate nucleotide hydrolase) . Proteomic comparisons revealed of 281 up-regulated and 96 down-regulated proteins in the ∆ ung ∆ dut cells as compared to the wild-type, many of them involved in nucleotide metabolism. Unexpectedly, genome uracylation did not change much the gross environmental endurance profile of P. putida , increased spontaneous mutagenesis by only 2-fold and supported expression of heterologous proteins well. Since U-enriched DNA is potentially degraded by the base excision repair of recipients encoding an uracil DNA glycosylase, we then tested the spread potential of genetic material originated in the ∆ ung ∆ dut cells either within the same species or to a commonly used E. coli strain. Transformation and conjugation experiments revealed that horizontal gene transfer of U-containing plasmids fared worse than those made of standard DNA by 2 orders of magnitude. While this figure does not guarantee certainty of containment, it suggests a general strategy for curbing dispersal of recombinant genetic constructs.
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Source: Chembiochem - Category: Biochemistry Authors: Algar E, Al-Ramahi Y, de Lorenzo V, Martínez-García E Tags: Chembiochem Source Type: research