Production of a recombinant antimicrobial peptide in transgenic plants using a modified VMA intein expression system
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- 作者：Morassutti ; Carla ; De Amicis ; Francesca ; Skerlavaj ; Barbara ; Zanetti ; Margherita ; Marchetti ; Stefano
- 关键词：Antimicrobial peptide ; Cathelicidin ; Intein ; Expression system ; Transgenic plant ; Biotherapeutic ; AAU ; acid-acetic-urea ; BTP ; bis-tris-propane ; DTT ; 1 ; 4-dithiothreitol ; ELISA ; enzyme-linked immunosorbent assay ; LB ; Luria&ndash ; Bertani ; MW ; molecular weigh
- 刊名：FEBS Letters
- 出版年：2002
- 出版时间：May 22, 2002
- 年：2002
- 卷：519
- 期：1-3
- 页码：141-146
- 全文大小：262 K
文摘
Tobacco plants were engineered to express SMAP-29, a mammalian antimicrobial peptide of innate immunity, as fusion protein with modified vacuolar membrane ATPase intein. The peptide was purified taking advantage of the intein-mediated self-cleaving mechanism. SMAP-29 was immunologically detected in the chromatographic eluate and appeared tightly bound to copurified plant proteins. Electrophoretic separation under disaggregating conditions indicated that the recombinant peptide was cleaved off by intein at the expected site and an overlay gel assay demonstrated that the peptide retained antimicrobial activity. These results indicate that a modified intein expression system can be used to produce pharmaceutical peptides in transgenic plants.