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.
journal_name
Appl Biochem Biotechnoljournal_title
Applied biochemistry and biotechnologyauthors
Kodrík D,Kludkiewicz B,Navrátil O,Skoková Habuštová O,Horáčková V,Svobodová Z,Vinokurov KS,Sehnal Fdoi
10.1007/s12010-013-0325-9subject
Has Abstractpub_date
2013-09-01 00:00:00pages
209-24issue
1eissn
0273-2289issn
1559-0291journal_volume
171pub_type
杂志文章abstract：:Sorghum was pretreated by sole ultrasound or supercritical carbon dioxide (scCO2), as well as the method combining both to intensify enzymatic hydrolysis. The effect of the time (1-5 h) and temperature (30-70 °C) on ultrasonic pretreatment was investigated, and the best condition was determined as 5 h and 50 °C with t...
journal_title：Applied biochemistry and biotechnology
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