Expression of CTB-proinsulin in transgenic chloroplasts

Abstract

Diabetes mellitus is presently recognized as the sixth leading cause of death in the United States, affecting over 20 million people. Diabetes is a condition characterized by high blood glucose due to an insulin deficiency or resistance. Type I, which comprises 5-10% of all cases, results from the destruction of pancreatic beta cells in the islets of Langerhans. The current treatment for type I diabetes is insulin administration through injection or pump. Purification, production, and storage of this insulin proves to be quite costly. By producing biologically functional insulin with oral delivery capabilities through chloroplast genetic engineering, many of these costs could be cut back. In addition, the possibility of providing the C-peptide currently lacking in commercially available insulin becomes available. The Daniell lab inserted cholera toxin B-subunit (CTB) fused proinsulin, containing three furin cleavage sites, (CTB-pins Fx3) into the tobacco chloroplasts of Petit Havana via particle gun bombardment. The insertion of three furin cleavage sites along with fusion to CTB will enable the insulin to sustain function when orally administered. Transgenic plants from the second generation were then analyzed and tested for quantification of the CTB-pins Fx3 gene. Tobacco leaves of varying ages were compared to determine expression levels. The resulting data is pertinent for future production of both orally deliverable insulin and more cost effective injectable insulin.

Notes

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Thesis Completion

2008

Semester

Fall

Advisor

Daniell, Henry

Degree

Bachelor of Science (B.S.)

College

Burnett College of Biomedical Sciences

Degree Program

Molecular Biology and Microbiology

Subjects

Biomedical Sciences -- Dissertations, Academic;Dissertations, Academic -- Biomedical Sciences

Format

Print

Identifier

DP0022252

Language

English

Access Status

Open Access

Length of Campus-only Access

None

Document Type

Honors in the Major Thesis

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