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J Appl Microbiol ; 109(5): 1845-52, 2010 Nov.
Article in English | MEDLINE | ID: mdl-20666867

ABSTRACT

AIM: To develop an applicable vector system and a transformation method for the manipulation of Dietzia spp. METHODS AND RESULTS: The pNV18 Nocardia-E. coli shuttle vector was tested and found to be a replicating plasmid in Dietzia sp. E1. With the use of pNV18, an electroporation method was optimized for the transformation of Dietzia sp. E1, and a transformation efficiency suitable for genetic manipulations was achieved (2·18×10(4) transformants µg(-1) DNA). The method was also applied for the transformation of Dietzia cinnamea, D. maris, D. natronolimnaea and D. psychralcaliphila. CONCLUSIONS: The first applicable vectors and a simple electroporation protocol enabling the manipulation of several Dietzia spp. are presented. SIGNIFICANCE AND IMPACT OF THE STUDY: Dietzia spp. have clinical, industrial and great environmental importance; however, the analysis of the Dietzia genus is currently hampered by the lack of manipulation techniques. The presented basic tools allow the genetic analysis of several Dietzia species, including the human disease-associated Dietzia maris.


Subject(s)
Actinomycetales/genetics , Biotechnology/methods , Electroporation/methods , Genetic Techniques , Actinomycetales/growth & development , Culture Media , Genetic Vectors/genetics , Plasmids/genetics
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