Development of the System of Plasmid Transfer to Strains Streptomyces avermitilis ATCC 31272 andSaccharopolyspora erythraea ATCC 11635 by the Method of Intergeneric Conjugation

Development of the System of Plasmid Transfer to Strains Streptomyces avermitilis ATCC 31272... A new method of plasmid DNA transfer from the donor strainEscherichia coliS17-1 to the erythromycin-producing strainSaccharopolyspora erythraeaand avermectin-producing strain Streptomyces avermitilis via intergeneric conjugation was proposed. The optimal parameters of the method were chosen for increasing the efficiency of crosses and ensuring easily reproducible results. The behavior of the multicopy plasmid pPM803 and the integration vector pTO1 along with a number of new plasmids specially created by us, was examined in these strains. A new plasmid vector (pSI60) capable of integrating into the chromosome of actinomycetes at the integration site of the temperate actinophage φC31 was constructed. This vector possesses unique sites convenient for cloning and may be stably maintained in exconjugants of S. avermitilis and in the model strain Streptomyces lividans.The gene encoding resistance to spectinomycin and streptomycin was cloned into the vector pSI60 in this strain. For cloning in strain Sac. erythraea, vectors pSI261–280, which integrate into the chromosome via homology with the cloned DNA and can be stably maintained in exconjugants, were constructed. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Russian Journal of Genetics Springer Journals

Development of the System of Plasmid Transfer to Strains Streptomyces avermitilis ATCC 31272 andSaccharopolyspora erythraea ATCC 11635 by the Method of Intergeneric Conjugation

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Publisher
Kluwer Academic Publishers-Plenum Publishers
Copyright
Copyright © 2003 by MAIK “Nauka/Interperiodica”
Subject
Biomedicine; Human Genetics
ISSN
1022-7954
eISSN
1608-3369
D.O.I.
10.1023/A:1023783600929
Publisher site
See Article on Publisher Site

Abstract

A new method of plasmid DNA transfer from the donor strainEscherichia coliS17-1 to the erythromycin-producing strainSaccharopolyspora erythraeaand avermectin-producing strain Streptomyces avermitilis via intergeneric conjugation was proposed. The optimal parameters of the method were chosen for increasing the efficiency of crosses and ensuring easily reproducible results. The behavior of the multicopy plasmid pPM803 and the integration vector pTO1 along with a number of new plasmids specially created by us, was examined in these strains. A new plasmid vector (pSI60) capable of integrating into the chromosome of actinomycetes at the integration site of the temperate actinophage φC31 was constructed. This vector possesses unique sites convenient for cloning and may be stably maintained in exconjugants of S. avermitilis and in the model strain Streptomyces lividans.The gene encoding resistance to spectinomycin and streptomycin was cloned into the vector pSI60 in this strain. For cloning in strain Sac. erythraea, vectors pSI261–280, which integrate into the chromosome via homology with the cloned DNA and can be stably maintained in exconjugants, were constructed.

Journal

Russian Journal of GeneticsSpringer Journals

Published: Oct 7, 2004

References

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