Expression and purification of recombinant forms of two Arabidopsis thaliana PR-10 homologues (MLP423 and Bet v 1)

  • Author / Creator
  • Pathogenesis related 10 (PR-10) proteins have important roles in mediating plant abiotic and biotic stress tolerance. Most of the members of this family possess ribonuclease activity. The homologues of PR-10 (ABR17) protein, MLP423 and Bet v 1, may have similar biological and physiological functions. The deduced amino acid sequence of MLP423 and Bet v 1, when compared to ABR17, indicated several conserved amino acids and a conserved P-loop motif (GxGGxGxxK). In the present investigation, cDNAs encoding Arabidopsis thaliana MLP423 and Bet v 1 were isolated and cloned, recombinant proteins expressed in E. coli, and purified using Ni-NTA and size exclusion chromatography. Both in-solution and in-gel RNase activity assays were conducted to determine whether these recombinant proteins possessed RNase activity, which indicated that neither MLP423 nor Bet v 1 possessed RNase activity. Our results are discussed in the context of the possible functions of these PR-10 homologues in plants.

  • Subjects / Keywords
  • Graduation date
    Fall 2013
  • Type of Item
  • Degree
    Master of Science
  • DOI
  • License
    This thesis is made available by the University of Alberta Libraries with permission of the copyright owner solely for non-commercial purposes. This thesis, or any portion thereof, may not otherwise be copied or reproduced without the written consent of the copyright owner, except to the extent permitted by Canadian copyright law.