Crude protein extraction protocol for phage N15 protelomerase in vitro enzymatic assays

Qingwen Chen, Kumaran Narayanan

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    3 Citations (Scopus)

    Abstract

    The phage N15 protelomerase enzyme (TelN) is essential for the replication of its genome by resolution of its telRL domain, located within a telomerase occupancy site (tos), into hairpin telomeres. Isolation of TeIN for in vitro processing of tos, however, is a highly complex process, requiring multiple purification steps. In this study a simplified protocol for crude total protein extraction is described that retains the tos-cleaving activity of TeIN for at least 4 weeks, greatly simplifying in vitro testing of its activity. This protocol may be extended for functional analysis of other phage and bacterial proteins, particularly DNA-processing enzymes
    Original languageEnglish
    Pages (from-to)169 - 171
    Number of pages3
    JournalAnalytical Biochemistry
    Volume414
    Issue number1
    DOIs
    Publication statusPublished - 2011

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