Factors affecting recombinant frequency in protoplast fusions of Streptomyces coelicolor.
David A. Hopwood,Helen M. Wright +1 more
TLDR
The frequency of recombinants in the progeny could be significantly enhanced by ultraviolet irradiation of the parental protoplast suspensions immediately before fusion.Abstract:
Summary: The optimum concentration of polyethylene glycol 1000 (PEG) for the production of recombinants through protoplast fusion in Streptomyces coelicolor was about 50% (w/v). The addition of 14% (v/v) dimethyl sulphoxide to the fusion mixture enhanced recombination frequencies, but only at sub-optimal PEG concentrations. After treatment of protoplasts with 50% PEG for 1 min, the frequency of recombinants in a multi-factor ‘cross’ sometimes exceeded 20% of the total progeny. The frequency of recombinants in the progeny could be significantly enhanced by ultraviolet irradiation of the parental protoplast suspensions immediately before fusion.read more
Citations
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Journal ArticleDOI
Genome shuffling leads to rapid phenotypic improvement in bacteria
Ying-Xin Zhang,Kim Perry,Victor A. Vinci,Keith A. Powell,Willem P. C. Stemmer,Stephen B. del Cardayre +5 more
TL;DR: The whole-genome shuffling approach as discussed by the authors combines the advantage of multi-parental crossing allowed by DNA shuffling with the recombination of entire genomes normally associated with conventional breeding.
Patent
Evolution of whole cells and organisms by recursive sequence recombination
Stephen B. del Cardayre,Matthew Tobin,Willem P. C. Stemmer,Jon E. Ness,Jeremy Minshull,Phillip A. Patten,Venkiteswaran Subramanian,Linda A. Castle,Claus M. Krebber,Steven H. Bass,Ying-Xin Zhang,Tony Cox,Gjalt W. Huisman,Ling Yuan,Joseph A. Affholter +14 more
TL;DR: In this paper, the authors employ iterative cycles of recombination and selection/screening for evolution of whole cells and organisms toward acquisition of desired properties such as enhanced recombinogenicity, genome copy number, and capacity for expression and/or secretion of proteins and secondary metabolites.
Book ChapterDOI
Genetics of Industrial Microorganisms
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TL;DR: Antibiotic-blocked mutants have been used in several recent biosynthetic studies and their utility in the synthesis of new antibiotics via interspecific matings is discussed, and gene-transfer in microorganisms is discussed.
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Genome shuffling: Progress and applications for phenotype improvement
TL;DR: The advantage of genome shuffling is presented, the procedure of this technology is introduced, the applications of this approach for phenotype improvement are summarized, and perspective on the development of this method in the future is given.
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Genome-shuffling improved acid tolerance and l-lactic acid volumetric productivity in Lactobacillus rhamnosus
TL;DR: Improved the acid tolerance and volumetric productivity of an industrial strain Lactobacillus rhamnosus ATCC 11443 by genome shuffling.
References
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Journal ArticleDOI
Formation and Reversion of Streptomycete Protoplasts: Cultural Condition and Morphological Study
M. Okanishi,K. Suzuki,H. Umezawa +2 more
TL;DR: Combinations of appropriate concentrations of MgCl2 and CaCl2 were significantly effective in the reversion as well as in the stability of protoplasts.
Journal ArticleDOI
Transformation of plasmid DNA into Streptomyces at high frequency
TL;DR: The development of a plasmid transformation system for Streptomyces which should allow the cloning of any DNA sequence into these organisms and which, potentially, provides a means of directly manipulating the pathways of antibiotic production.
Journal ArticleDOI
Genetic recombination through protoplast fusion in Streptomyces.
TL;DR: Recombination frequencies achieved by this technique are so high that selectable markers could be dispensed with, at least in certain kinds of strain improvement programmes, and the availability of a simple generally applicable procedure to recombine actinomycete strains at high frequency is reported.
Journal ArticleDOI
Genetic Recombination in Streptomyces fradiae by Protoplast Fusion and Cell Regeneration
TL;DR: Fusion of protoplasts carrying different auxotrophic or chromosomal drug-resistance markers was achieved by treatment with polyethylene glycol, and high frequencies of stable genetic recombinants were obtained.
Journal ArticleDOI
Bacterial protoplast fusion: Recombination in fused protoplasts of Streptomyces coelicolor
David A. Hopwood,Helen M. Wright +1 more
TL;DR: The generation of nearly random populations of recombinants between two or more parent strains by protoplast fusion under the conditions described appears to have simple applications in industrial and academic strain construction.
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Genetic recombination through protoplast fusion in Streptomyces.
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