Multiplex PCR assay for detection of recombinant genes encoding fatty acid desaturases fused with lichenase reporter protein in GM plants

Thermostable lichenase encoded by licB gene of Clostridium thermocellum can be used as a reporter protein in plant, bacterial, yeast, and mammalian cells. It has important advantages of high sensitivity and specificity in qualitative and quantitative assays. Deletion variants of LicB (e.g., LicBM3) retain its enzymatic activity and thermostability and can be expressed in translational fusion with target proteins without compromising with their properties. Fusion with the lichenase reporter is especially convenient for the heterologous expression of proteins whose analysis is difficult or compromised by host enzyme activities, as it is in case of fatty acid desaturases occurring in all groups of organisms. Recombinant desaturase-lichenase genes can be used for creating genetically modified (GM) plants with improved chill tolerance. Development of an analytical method for detection of fused desaturase-lichenase transgenes is necessary both for production of GM plants and for their certification. Here, we report a multiplex polymerase chain reaction method for detection of desA and desC desaturase genes of cyanobacteria Synechocystis sp. PCC6803 and Synechococcus vulcanus, respectively, fused to licBM3 reporter in GM plants.     

Selection of Optimal Parameters of Multilayer Plates at Nonstationary Loading

This investigation features two approaches to optimal design of multilayer plates at impulse loading. The first approach deals with minimization of stresses in the plate layers by selecting the elastic properties of the fillers. The second approach is based on a twolevel synthesis procedure applied to the problem of optimal design of multilayer plates with a minimum mass. The effectiveness of these approaches is illustrated by numerical examples.Sommario. L'analisi è tesa alla caratterizzazione di due procedure di progettazione ottimale di piastre multistrato ad azioni impulsive. La prima procedura concerne con la minimizzazione dello sforzo negli strati della piastra selezionando le proprietà elastiche dei riempimenti.II secondo approccio e basato su di una procedura a due livelli di sintesi applicata al problema del disegno ottimo di piastre multistrato con una massa minima. L'efficacia di questi approcci e illustrata tramite esempi numerici.