Date of Award
10-3-2019
Document Type
Thesis
Degree Name
Biology, MS
First Advisor
Maureen Dolan
Second Advisor
Jianfeng Xu
Committee Members
Argelia Lorence; Lorin Neuman-Lee
Call Number
LD 251 .A566t 2019 C36
Abstract
Despite the advantages of plant-produced recombinant proteins, the most important bottleneck that limits the commercialization is the low protein yields. Plants have a unique type of O-glycosylation that has potential to enhance the stability and solubility of recombinant proteins. Specifically, target gene sequences are fused with a sequence to code for hydroxyproline-O-glycosylated peptide (HypGP) tags. We targeted the expression of a very unstable rainbow trout interleukin 22 (rtIL-22) and a readily-monitored enhanced green fluorescent protein (EGFP). The HypGP-tagged rIL-22 accumulation in planta was significantly enhanced over the untagged protein and showed increase stability. Changes in gene expression of the marker genes indicated bioactivity of HypGP-tagged rtIL-22. Additionally, we explored the impact of water deficit on Nicotiana benthamiana for increasing expression and recovery of the HypGP-tagged EGFP. Finally, an apoplast wash fluid (AWF) extraction method was optimized to determine the spatial location of different HypGP-tagged proteins in the whole plant system.
Rights Management
This work is licensed under a Creative Commons Attribution 4.0 International License.
Recommended Citation
Calvo, Cristofer, "Optimizing Hydroxyproline-O-Glycan Engineering in Tobacco Transient Protein Expression: Fish IL-22 And Enhanced Green Fluorescent Protein (EGFP)" (2019). Student Theses and Dissertations. 396.
https://arch.astate.edu/all-etd/396
