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Expression of Arabidopsis thaliana cellulose synthase proteins and associated proteins in a Spodoptera frugiperda cell line

Understanding how cellulose synthesis occurs is key to understanding the
formation of the plant cell wall. This understanding could also be key to modifying
cellulose production to permit more efficient extraction of glucose from cellulose for the
production of biobased materials. Cellulose biosynthesis is carried out by cellulose
synthases; transmembrane multimeric processive glycosyltransferases responsible for
polymerizing UDP‐glucose into glucan chains. Thirty‐six glucan chains bind together in
parallel to form elementary cellulose microfibrils. Due to the essential nature of
cellulose synthases for plant survival and the recalcitrant nature of the cell wall to
chemical and enzymatic digestion, the cellulose synthases can be very difficult to
analyze by traditional approaches. In an attempt to circumvent some of the issues of
studying cellulose synthases, the cellulose synthase genes CESA1 and CESA3, along with
the cell wall associated genes COBRA, DET3 and POM1 were recombined into an
engineered Autographa californica nucleopolyhedron virus and expressed in Spodoptera
fruigiperda ovarian cells. Although recombinant protein could be detected for CESA1
and CESA3, C14‐glucose incorporation on baculovirus infected cell lines have given
inconclusive results to the cellulose synthase activity of the CESA1 and CESA3 proteins.
With further optimization of the protein expression of CESA1 and optimization of the
variability in the C14‐glucose incorporation assays, the baculovirus system may prove a
useful tool for studying the cellulose synthases. / UOIT

Identiferoai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:OOSHDU.10155/274
Date01 October 2012
CreatorsLyons, Jessy
ContributorsBonetta, Dario
Source SetsLibrary and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada
LanguageEnglish
Detected LanguageEnglish
TypeThesis

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