Production of a recombinant antimicrobial peptide in transgenic plants using a modified VMA intein expression system

FEBS Letters

Zanetti, Margherita²  Marchetti, Stefano¹  Morassutti, Carla¹  De Amicis, Francesca¹  Skerlavaj, Barbara² 

[1] DPVTA, Università di Udine, Via delle Scienze 208, 33100 Udine, Italy;DSTB, Università di Udine, Piazzale Kolbe 4, 33100 Udine, Italy

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–Bertani;    MW;    molecular weight;    PAGE;    polyacrylamide gel electrophoresis;    SDS;    sodium dodecyl sulphate;    VMA;    vacuolar membrane ATPase;   

DOI  :  10.1016/S0014-5793(02)02741-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown