Methods for the preparation of ordered arrays of full-length proteins tagged with affinity labels for capture.

John Sutherland (Other)

    Research output: Patent

    Abstract

    The present invention describes methods for the prepn. of protein arrays of full length proteins. The method involves incorporating a specific label into each protein of a proteome at a position that does not affect protein structure or function. The protein can then be captured on a substrate via the label, e.g. by biotin-avidin or antibody-epitope interactions. The use of such arrays in screening methods is also described. An expression vector that creates a hexahistidine-tagged version of a protein and that uses a green fluorescent protein as reporter moiety to select for in-frame expression is described. Methods and primers for amplifying sequences from a cDNA library that avoid a bias towards stop codon-contg. sequences are also described. Reconstruction expts. in which specific activities were recovered from a small defined library are reported. [on SciFinder (R)]
    Original languageEnglish
    Patent numberPCT Int. Appl.
    Publication statusPublished - 2001

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