Morassutti Carla, De Amicis Francesca, Skerlavaj Barbara, Zanetti Margherita, Marchetti Stefano
DPVTA, Università di Udine, Via delle Scienze 208, Udine, Italy.
FEBS Lett. 2002 May 22;519(1-3):141-6. doi: 10.1016/s0014-5793(02)02741-2.
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.
烟草植株经过基因工程改造，以表达SMAP - 29（一种先天性免疫的哺乳动物抗菌肽），它作为与修饰的液泡膜ATP酶内含肽的融合蛋白。利用内含肽介导的自我切割机制对该肽进行了纯化。在色谱洗脱液中通过免疫检测到了SMAP - 29，并且它似乎与共纯化的植物蛋白紧密结合。在解离条件下的电泳分离表明，重组肽在预期位点被内含肽切割，并且覆盖胶试验表明该肽保留了抗菌活性。这些结果表明，一种修饰的内含肽表达系统可用于在转基因植物中生产药用肽。
