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Phenotypic expression of PCR-generated random mutations in a Pseudomonas putida gene after its introduction into an Acinetobacter chromosome by natural transformation

Article Abstract:

Localized sets of random point mutations generated by polymerase chain reaction amplification can be transferred to the chromosome of Acinetobacter ADP1 strain in an efficient manner using natural transformation. The natural transformation method eliminates the need for cloning of PCR fragments in plasmids since PCR-amplified DNA fragments are internalized by cells and directly introduced into their genomes by homologous recombination. Results show that homologous recombination of the PCR-amplified DNA fragments into a specific chromosomal docking site from which they can be expressed is made possible by nucleotide sequence identity between the flanking regions and corresponding chromosomal segments in a cloned Acinetobacter recipient.

Author: Ornston, L. Nicholas, Kok, Ruben G., Young, David M.
Publisher: American Society for Microbiology
Publication Name: Applied and Environmental Microbiology
Subject: Biological sciences
ISSN: 0099-2240
Year: 1999
Gene mutations, Gene mutation, Pseudomonas putida, Chromosomes, Gene amplification

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Development and use of a reverse transcription-PCR assay to study expression of Tri5 by Fusarium species in vitro and in planta

Article Abstract:

A reverse transcription-PCR assay was developed to investigate the expression of Tri5 by Fusarium species in planta and in vitro. An increase in Tri5 expression was observed following treatment of Fusarium culmorum with fungicides. An inverse relationship between Tri5 expression and biomass was also observed. Results provide an enhanced understanding of the influence of different host tissues and chemical control agents on trichothecene production.

Author: Doohan, F.M., Weston, G., Rezanoor, H.N., Parry, D.W., Nicholson, P.
Publisher: American Society for Microbiology
Publication Name: Applied and Environmental Microbiology
Subject: Biological sciences
ISSN: 0099-2240
Year: 1999
Reverse transcriptase, Fusarium, Microbiological assay

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Restoration of gene function by homologous recombination: From PCR to gene expression in one step

Article Abstract:

A simple method for single-step cloning of any PCR product into a plasmid is developed. A novel selection principle is applied, in which activation of a drug selection maker is achieved following homologous recombination.

Author: Yosef, Ido, Bloushtain, Noga, Shapira, Michal, Qimron, Udi
Publisher: American Society for Microbiology
Publication Name: Applied and Environmental Microbiology
Subject: Biological sciences
ISSN: 0099-2240
Year: 2004
Science & research

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Subjects list: Research, Genetic aspects, Polymerase chain reaction, Gene expression
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