Title page for ETD etd-04262000-17260046


Type of Document Master's Thesis
Author Kositsup, Boonthida
Author's Email Address bkositsu@vt.edu
URN etd-04262000-17260046
Title Tissue and Cell-Type Localization and Partial Characterization of a Xylem Papain-Type Cysteine Protease From Arabidopsis
Degree Master of Science
Department Horticulture
Advisory Committee
Advisor Name Title
Beers, Eric P. Committee Chair
Hess, John L. Committee Member
Welbaum, Gregory E. Committee Member
Keywords
  • programmed cell death
  • cysteine protease
  • Arabidopsis thaliana
  • tracheary element
  • xylem
  • localization
Date of Defense 2000-04-18
Availability unrestricted
Abstract
Cysteine proteases are associated with xylem tracheary element differentiation. XCP1 was recently identified as a xylem-specific cysteine protease in Arabidopsis (Zhao, et al., 2000). For this study a recombinant polyhistidine-tagged XCP1 (XCP1H6) was expressed and purified from an E. coli expression system. A polyclonal anti-XCP1 antibody was produced using purified XCP1H6. Immunoblot analysis of a developmental time course of xylem and bark protein extracted from root-hypocotyl segments demonstrated that XCP1 was expressed in xylem only. Further analysis under optimized immunoblot conditions, however, revealed that anti-XCP1 antibody reacted with protein present in both xylem and bark. The vast majority of immunoreactivity, however, was restricted to xylem. Cell-type localization of GUS expression under the control of a putative XCP1 promoter indicated that the XCP1 promoter specifies expression of XCP1 in tracheary elements in leaves, stems, roots and flowers. XCP1 promoter-driven GUS activity was not associated with senescing tissues.
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