Expression analysis of human -secretase in transgenic tomato fruits

Original Research Article

Protein Expression and Purification, Volume 82, Issue 1, March 2012, Pages 125-131 H.-S. Kim, J.-W. Youm, K.-B. Moon, J.-H. Ha, Y.-H. Kim, H. Joung, J.-H. Jeon

Abstract
An emerging strategy in biomanufacturing involves using transgenic plants to express recombinant pharmaceutical and industrial proteins in large quantities. -Site APP cleaving enzyme 1 (-secretase 1, BACE1) is an enzyme involved in the abnormal production of A42, the major component of senile plaques in Alzheimers disease (AD). Thus, BACE1 represents a key target protein in the development of new potential drugs to treat Alzheimers disease. We aimed to develop a tomato-derived recombinant BACE1 (rBACE1) protein to serve as a vaccine antigen that would promote an immune response. We utilized a plant expression cassette, pE8BACE, to optimize BACE1 expression in tomato fruits. Polyemerase chain reaction and Southern blot analyses verified integration of the BACE1 gene into the plant genome. Northern and Western blot analyses demonstrated successful mRNA and protein expression of rBACE1, respectively; the Sensizyme assay kit estimated the expression level of rBACE1 protein at 136 7 ng mg total soluble protein. The tomato-derived rBACE1 retains its activity for a long storage period at cool or room temperature, and is highly resistant to degradation in conditions such as low acidity. Tomato-derived rBACE1 was severely degraded by heat or boiling. The proteolytic activity of tomato-derived rBACE1, confirmed by fluorescence resonance transfer assay, was similar to that of a commercial sample of Escherichia coli-derived BACE1.
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Tomat tahan dingin, disisipkan gen ikan flonder Tanaman tahan kering, Nyamuk yang membawa tetrasilin, yang mebuat larva mati http://www.biosafety-info.net

Sebetulnya tujuan utama dari teknik breeding tradisional maupun bioteknologi modern sama saja, yaitu bagaimana caranya agar dapat menyisipkan sebuah gen atau beberapa gen dari sebuah organisme donor yang membawa atau memiliki sifat-sifat baik yang dikehendaki ke dalam suatu organisme yang tidak memiliki sifat-sifat (trait) tersebut. Lebih dari 50 jenis tanaman pangan GMO telah lolos dari uji dan review pemerintah federal AS dan sekitar 100 jenis komoditas GMO baru sedang mengalami uji lapang. Produk Jagung bt Problema Kematian pada hewan non target Solusi Pemaparan kondisi Penggunaan dari pestisida jua memberikan efek yang lebih buruk, bahkan secara langsung maupun tidak langsung maupun tidak pada lingkungan

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