Protease inhibitor from insect silk-activities of derivatives expressed in vitro and in transgenic potato.
Appl Biochem Biotechnol
; 171(1): 209-24, 2013 Sep.
Article
em En
| MEDLINE
| ID: mdl-23824530
ABSTRACT
Several recombinant derivatives of serine protease inhibitor called silk protease inhibitor 2 (SPI2), which is a silk component in Galleria mellonella (Lepidoptera, Insecta), were prepared in the expression vector Pichia pastoris. Both the native and the recombinant protease inhibitors were highly active against subtilisin and proteinase K. The synthetic SPI2 gene with Ala codon in the P1 position was fused with mGFP-5 to facilitate detection of the transgene and its protein product. A construct of the fusion gene with plant regulatory elements (promoter 35S and terminator OCS) was inserted into the binary vector pRD400. The final construct was introduced into Agrobacterium tumefaciens that was then used for genetic transformation of the potato variety Velox. The transgene expression was monitored with the aid of ELISA employing polyclonal antibody against natural SPI2. In vitro tests showed increased resistance to the late blight Phytophthora infestans in several transformed lines. No effect was seen on the growth, mortality, life span or reproduction of Spodoptera littoralis (Lepidoptera, Insecta) caterpillars, while feeding on transformed potato plants expressing the fusion protein, indicating that the transformed potatoes may be harmless to non-target organisms.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Solanum tuberosum
/
Engenharia Genética
/
Proteínas de Insetos
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Lepidópteros
Limite:
Animals
Idioma:
En
Revista:
Appl Biochem Biotechnol
Ano de publicação:
2013
Tipo de documento:
Article
País de afiliação:
República Tcheca