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Uridine phosphorylase from Shewanella oneidensis MR-1: Heterological expression, regulation, transcription, and properties

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Abstract

A DNA fragment containing a promoter-operator and structural parts of the uridine phosphorylase gene from Shewanella oneidensis MR-1 was cloned. Cross-heterological expression of the udp genes from Sh. oneidensis MR-1 and Escherichia coli under the control of authentic regulatory regions is shown. The UDP protein accumulates in an active form in the cytoplasmic fraction of cells. The recombinant UDP protein from Sh. oneidensis MR-1 obtained by heterological expression was isolated and characterized. E. coli udp gene promoter activity was observed during heterological expression in Sh. oneidensis MR-1 cells under both aerobic and anaerobic conditions.

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Abbreviations

OD:

optical density

PAAG:

polyacrylamide gel

PCR:

polymerase chain reaction

LB medium:

Luria-Bertani medium

dNTP:

deoxynucleoside triphosphate(s)

TSB:

tryptone soy broth

UDP:

uridine phosphorylase

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Authors and Affiliations

  1. State Research Institute of Genetics and Selection of Industrial Microorganisms (GosNIIgenetika), Moscow, 117545, Russia

    N. N. Mordkovich, V. A. Manuvera, V. P. Veiko & V. G. Debabov

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  1. N. N. Mordkovich
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  2. V. A. Manuvera
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  3. V. P. Veiko
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  4. V. G. Debabov
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Correspondence to N. N. Mordkovich.

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Original Russian Text © N.N. Mordkovich, V.A. Manuvera, V.P. Veiko, V.G. Debabov, 2012, published in Biotekhnologiya, 2012, No. 1, pp. 21–30.

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Mordkovich, N.N., Manuvera, V.A., Veiko, V.P. et al. Uridine phosphorylase from Shewanella oneidensis MR-1: Heterological expression, regulation, transcription, and properties. Appl Biochem Microbiol 48, 716–722 (2012). https://doi.org/10.1134/S0003683812090037

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  • DOI: https://doi.org/10.1134/S0003683812090037

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