Recurrent Selection for Transgene Activity Levels in Maize Results in Proxy Selection for a Native Gene with the Same Promoter

High activity levels of a transgene can be very useful, making a transgene easier to evaluate for safety and efficacy. High activity levels can also increase the economic benefit of the production of high value proteins in transgenic plants. The goal of this research is to determine if recurrent selection for activity of a transgene will result in higher activity, and if selection for activity of a transgene controlled by a native promoter will also increase protein levels of the native gene with the same promoter. To accomplish this goal we used transgenic maize containing a construct encoding green fluorescent protein controlled by the promoter for the maize endosperm-specific 27kDa gamma zein seed storage protein. We carried out recurrent selection for fluorescence intensity in two breeding populations. After three generations of selection, both selected populations were significantly more fluorescent and had significantly higher levels of 27kDa gamma zein than the unselected control populations. These higher levels of the 27kDa gamma zein occurred independently of the presence of the transgene. The results show that recurrent selection can be used to increase activity of a transgene and that selection for a transgene controlled by a native promoter can increase protein levels of the native gene with the same promoter via proxy selection. Moreover, the increase in native gene protein level is maintained in the absence of the transgene, demonstrating that proxy selection can be used to produce non-transgenic plants with desired changes in gene expression

Experimental Research on the Development of the Composition of the Transdermal Therapeutic System of Anti-inflammatory Action Based on Composition of Natural Substances

The aim of the work is to develop the composition of the transdermal therapeutic system of anti-inflammatory action based on active pharmaceutical ingredients of natural origin. Materials and methods. Selection of adhesive materials, plasticizers and solvents, the effect of introduction to the base of active pharmaceutical ingredients, determination of optimal temperature and drying time, pharmacological screening was performed by physicochemical, biopharmaceutical and biological research methods. Results. The composition of the hydrophilic adhesive composition, which includes polyvinylpyrrolidone, eudragit (adhesives), PEO-400 (plasticizer), isopropyl alcohol (solvent) 50: 8: 12: 30, respectively. This composition has excellent organoleptic properties, is easily applied to the polymer base, holds its shape well (does not flow beyond the base), is applied in a thin layer and evenly distributed on the base. The optimal mode of drying of the adhesive composition based on PVP is 75 °C for 30 min, based on PVA – 75 °C for 35 min, the composition containing eudragit - 50 °C for 10 min. It is established that the addition of active substances to the adhesive composition does not adversely affect its properties. The study of antiexudative activity using different combinations of substances of natural origin as API and found that the best anti-inflammatory properties (25 %) have a sample No. 4, which includes dry extracts of white willow bark and sage leaves and quercetin 3:1:3 in accordance. Conclusions. Based on physicochemical, biopharmaceutical and pharmacological studies, the composition of the transdermal therapeutic system of anti-inflammatory action with active pharmaceutical ingredients of natural origin has been develope

Experimental Research on the Development of the Composition of the Transdermal Therapeutic System of Anti-inflammatory Action Based on Composition of Natural Substances

The aim of the work is to develop the composition of the transdermal therapeutic system of anti-inflammatory action based on active pharmaceutical ingredients of natural origin. Materials and methods. Selection of adhesive materials, plasticizers and solvents, the effect of introduction to the base of active pharmaceutical ingredients, determination of optimal temperature and drying time, pharmacological screening was performed by physicochemical, biopharmaceutical and biological research methods. Results. The composition of the hydrophilic adhesive composition, which includes polyvinylpyrrolidone, eudragit (adhesives), PEO-400 (plasticizer), isopropyl alcohol (solvent) 50: 8: 12: 30, respectively. This composition has excellent organoleptic properties, is easily applied to the polymer base, holds its shape well (does not flow beyond the base), is applied in a thin layer and evenly distributed on the base. The optimal mode of drying of the adhesive composition based on PVP is 75 °C for 30 min, based on PVA – 75 °C for 35 min, the composition containing eudragit - 50 °C for 10 min. It is established that the addition of active substances to the adhesive composition does not adversely affect its properties. The study of antiexudative activity using different combinations of substances of natural origin as API and found that the best anti-inflammatory properties (25 %) have a sample No. 4, which includes dry extracts of white willow bark and sage leaves and quercetin 3:1:3 in accordance. Conclusions. Based on physicochemical, biopharmaceutical and pharmacological studies, the composition of the transdermal therapeutic system of anti-inflammatory action with active pharmaceutical ingredients of natural origin has been develope

Effect of selection on germination rate.

<p>Data points are mean number of plants per row values versus generation, shown with the linear regression trend lines. The equations of the lines for the selected and unselected populations are y = -0.15x + 22.45 (R² = 0.1579) and y = -0.225x + 23.133 (R² = 0.4023) respectively.</p

Analysis of variance of fluorescence, germination rate, and seed mass.

<p>Analysis of variance of fluorescence, germination rate, and seed mass.</p

Maize segregating for various levels of GFP activity.

<p>The top panel is in white light and the bottom panel in blue light (485nm) with an orange filter (535nm).</p