NDLI: Improved in Planta Expression of the Human Islet Autoantigen Glutamic Acid Decarboxylase (GAD65)

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Tritrophic Interactions Between Transgenic Potato Expressing Snowdrop Lectin (GNA), an Aphid Pest (Peach–Potato Aphid; Myzus Persicae (Sulz.) and a Beneficial Predator (2-Spot Ladybird; Adalia Bipunctata L.)

Transfer and Expression of an Artificial Storage Protein (ASP1) Gene in Cassava (Manihot Esculenta Crantz)

Simple Databases to Monitor the Generation and Organisation of Transgenic Mouse Colonies

Transgenic Research : Volume 12, Issue 1, February 2003

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Transgenic Research : Volume 10

Transgenic Research : Volume 9

Transgenic Research : Volume 8

Transgenic Research : Volume 7

Transgenic Research : Volume 6

Improved in Planta Expression of the Human Islet Autoantigen Glutamic Acid Decarboxylase (GAD65)

Content Provider	Springer Nature Link
Author	Avesani, Linda Falorni, Alberto Tornielli, Giovanni Battista Marusic, Carla Porceddu, Andrea Polverari, Annalisa Faleri, Claudia Calcinaro, Filippo Pezzotti, Mario
Copyright Year	2003
Abstract	The smaller isoform of the enzyme glutamic acid decarboxylase (GAD65) is a major islet autoantigen in autoimmune type 1 diabetes mellitus (T1DM). Transgenic plants expressing human GAD65 (hGAD65) are a potential means of direct oral administration of the islet autoantigen in order to induce tolerance and prevent clinical onset of disease. We have previously reported the successful generation of transgenic tobacco and carrot that express immunoreactive, full-length hGAD65. In the present study, we tested the hypothesis that the expression levels of recombinant hGAD65 in transgenic plants can be increased by targeting the enzyme to the plant cell cytosol and by mediating expression through the potato virus X (PVX) vector. By substituting the NH$_{2}$-terminal region of hGAD65 with a homologous region of rat GAD67, a chimeric GAD67$_{1-87}$/GAD65$_{88-585}$ molecule was expressed in transgenic tobacco plants. Immunolocalization analysis showed that immunoreactive GAD67/65 was found in the plant cell cytosol. By using a radio-immuno assay with human serum from a GAD65 autoantibody-positive T1DM patient, the highest expression level of the recombinant GAD67/65 protein was estimated to be 0.19% of the total soluble protein, compared to only 0.04% of wild-type hGAD65. Transient expression of wild-type, full-length hGAD65 in N. benthamiana mediated by PVX infection was associated with expression levels of immunoreactive protein as high as 2.2% of total soluble protein. This substantial improvement of the expression of hGAD65 in plants paves the way for immunoprevention studies of oral administration of GAD65-containing transgenic plant material in animal models of spontaneous autoimmune diabetes.
Starting Page	203
Ending Page	212
Page Count	10
File Format	PDF
ISSN	09628819
Journal	Transgenic Research
Volume Number	12
Issue Number	2
e-ISSN	15739368
Language	English
Publisher	Kluwer Academic Publishers
Publisher Date	2003-01-01
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Human Genetics Plant Sciences Animal Anatomy / Morphology / Histology
Content Type	Text
Resource Type	Article
Subject	Genetics Animal Science and Zoology Biotechnology Agronomy and Crop Science

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