Počet záznamů: 1

A Cost-effective Amino-acid-type Selective Isotope Labeling of Proteins Expressed in Leishmania tarentolae

  1. 1.
    0334906 - BC-A 2010 RIV US eng J - Článek v odborném periodiku
    Foldynová-Trantírková, Silvie - Matulová, J. - Dötsch, V. - Löhr, F. - Cirstea, I. - Alexandov, K. - Breitling, R. - Lukeš, Julius - Trantírek, Lukáš
    A Cost-effective Amino-acid-type Selective Isotope Labeling of Proteins Expressed in Leishmania tarentolae.
    Journal of Biomolecular Structure & Dynamics. Roč. 26, č. 6 (2009), s. 755-761 ISSN 0739-1102
    Grant CEP: GA ČR GP204/08/P585; GA AV ČR 1QS600220554; GA AV ČR KAN200100801; GA MŠk 2B06129
    Výzkumný záměr: CEZ:AV0Z60220518
    Klíčová slova: NMR * isotope labeling * protein expression * Leishmania * low-level enrichment
    Kód oboru RIV: EB - Genetika a molekulární biologie
    Impakt faktor: 1.124, rok: 2009

    We report a cost efficient approach for amino-acid-type selective isotope labeling of proteins expressed in Leishmania tarentolae. The method provides an economically advantageous alternative to recently established protocol for isotopic labeling using expensive synthetic media. The method is based on cultivation of the L. tarentolae expression strain in a cheap complex medium supplemented with labeled amino acid(s). In this protocol, a labeled amino acid is deliberately diluted in the medium of undefined composition, which leads to alow-level isotope enrichment upon protein over-expression. The economic advantage of the protocol is achieved by avoiding large volumes of expensive synthetic medium. Decreased sensitivity of a NMR experiment due to low-level isotope enrichment is compensated by a five- to seven-fold increase of the yield of the recombinant protein in complex mediumas compared to that in the synthetic medium.
    Trvalý link: http://hdl.handle.net/11104/0179515