Anal Biochem. 2009 Dec 1;395(1):77-85. Epub 2009 Jul 30.

Purification of the large ribosomal subunit via its association with the small subunit.

Simons SP, McLellan TJ, Aeed PA, Zaniewski RP, Desbonnet CR, Wondrack LM, Marr ES, Subashi TA, Dougherty TJ, Xu Z, Wang IK, LeMotte PK, Maguire BA.

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Department of Exploratory Medicinal Sciences, Pfizer Global Research and Development, Groton, CT 06340, USA.

Abstract

We have developed an affinity purification of the large ribosomal subunit from Deinococcus radiodurans that exploits its association with FLAG-tagged 30S subunits. Thus, capture is indirect so that no modification of the 50S is required and elution is achieved under mild conditions (low magnesium) that disrupt the association, avoiding the addition of competitor ligands or coelution of common contaminants. Efficient purification of highly pure 50S is achieved, and the chromatography simultaneously sorts the 50S into three classes according to their association status (unassociated, loosely associated, or tightly associated), improving homogeneity.

PMID:: 19646947; [PubMed - indexed for MEDLINE]

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Purification of the large ribosomal subunit via its association with the small subunit.

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