Plastid transformation for Rubisco engineering and protocols for assessing expression

Spencer M. Whitney, Robert E. Sharwood

    Research output: Chapter in Book / Conference PaperChapter

    12 Citations (Scopus)

    Abstract

    The assimilation of CO2 within chloroplasts is catalyzed by the bi-functional enzyme ribulose-1, 5-bisphosphate carboxylase/oxygenase, Rubisco. Within higher plants the Rubisco large subunit gene, rbcL, is encoded in the plastid genome, while the Rubisco small subunit gene, RbcS is coded in the nucleus by a multi-gene family. Rubisco is considered a poor catalyst due to its slow turnover rate and its additional fixation of O2 that can result in wasteful loss of carbon through the energy requiring photorespiratory cycle. Improving the carboxylation efficiency and CO2/O2 selectivity of Rubisco within higher plants has been a long-term goal which has been greatly advanced in recent times using plastid transformation techniques. Here we present experimental methodologies for efficiently engineering Rubisco in the plastids of a tobacco master-line and analyzing leaf Rubisco content.
    Original languageEnglish
    Title of host publicationChloroplast Biotechnology: Methods and Protocols
    EditorsPal Maliga
    Place of PublicationU.S.
    PublisherHumana Press
    Pages245-262
    Number of pages18
    ISBN (Electronic)9781627039956
    ISBN (Print)9781627039949
    DOIs
    Publication statusPublished - 2014

    Keywords

    • Rubisco enzymes
    • immuno-detection
    • tobacco

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