Production of a recombinant antimicrobial peptide in transgenic plants using a modified VMA intein expression system
Production of a recombinant antimicrobial peptide in transgenic plants using a modified VMA intein expression system
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DOI：
10.1016/s0014-5793(02)02741-2
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发表时间：
2002-05
期刊：
影响因子：
3.5
通讯作者：
C. Morassutti;F. De Amicis;B. Skerlavaj;M. Zanetti;S. Marchetti
中科院分区：
文献类型：
--
作者：
C. Morassutti;F. De Amicis;B. Skerlavaj;M. Zanetti;S. Marchetti
利用基因工程技术，将具有天然免疫功能的哺乳动物抗菌肽SMAP-29与修饰的液泡膜ATPase内含子融合表达。利用内含素介导的自切机制对多肽进行了纯化。在层析洗脱液中免疫检测到SMAP-29，并与接合的植物蛋白紧密结合。在解聚条件下的电泳分离表明，重组多肽被内含素在预期的位置切割，重叠凝胶实验表明该重组多肽仍具有抗菌活性。这些结果表明，改良的内含素表达系统可用于在转基因植物中生产药用多肽。
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.