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Title 

High-level expression of a human β-site APP cleaving enzyme in transgenic tobacco chloroplasts and its immunogenicity in mice

Authors 

JungWon YoumJae Heung JeonH KimSung Ran MinM S KimHyouk JoungWon Joong JeongHyun Soon Kim

Publisher 

Springer Verlag (Germany)

Issue Date 

2010

Citation 

Transgenic Research, vol. 19, no. 6, pp. 1099-1108

Keywords 

β-secretaseAlzheimer diseaseChloroplast transformationNicotiana tabacum L.Plant-derived vaccine

Abstract 

Plastid transformation has to date been applied to the expression of heterologous genes involved in agronomic traits and to the production of useful recombinant proteins. Here, we report a feasibility study for producing the human β-site APP cleaving enzyme (BACE) via transformation of tobacco chloroplasts. Stable integration of human BACE into the plastome was confirmed by PCR. Genomic Southern blot analysis detected the presence of the tobacco aadA and human BACE genes between trnI and trnA in the plastome. Northern blot analysis revealed that the aadA and BACE genes were both properly transcribed into a dicistronic transcriptional unit. Human BACE protein expression in transplastomic tobacco was determined by western blot analysis. ELISA analysis revealed that, based on a dilution series of E. coli-derived BACE as a standard, transplastomic lines accumulated BACE to levels of 2. 0% of total soluble proteins. When mice were gavaged with the transplastomic tobacco extracts, they showed an immune response against the BACE antigen. The successful production of plastid-based BACE protein has the potential for developing a plant-based vaccine against Alzheimer disease.

ISSN 

0962-8819

Link 

http://dx.doi.org/10.1007/s11248-010-9383-8

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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