ABSTRACT
A set of module vectors for stable or transient gene expression in plants was constructed with regard to the majority of factors ensuring efficient heterologous gene expression in plants. The vectors are convenient to clone new regulatory elements and genes of interest via simple molecular cloning procedures. The vectors can be used to obtain transgenic plants with stable heterologous gene expression as well as to achieve transient expression because one vector includes the gene for the tomato bushy stunt virus p19 protein, which acts as a suppressor of posttranscriptional gene silencing.
Subject(s)
Gene Expression Regulation, Plant , Genetic Vectors , Plants, Genetically Modified , Base Sequence , Genes, Reporter , Molecular Sequence Data , Plasmids/genetics , RNA Interference , Nicotiana/genetics , Viral Proteins/geneticsABSTRACT
The basic methods used in current practice for stable and transient expression of heterologous genes in plants are presented and compared. The key areas of research in the heterologous expression of genes in plants have been identified by analyzing literature and experimental data: modeling of metabolic pathways; creation of marker-free transgenic plants; the search for new regulatory elements and plant genes influencing the efficiency of expression of heterologous genes in plants; development of new methods for analyzing of transgenic plants and new approaches to the expression of heterologous genes in plants.
Subject(s)
Gene Expression , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Genetic MarkersABSTRACT
Lysophosphatidic acid (LPA) is a lipid mediator required for maintaining homeostasis of numerous physiological functions and also involved in development of some pathological processes through interactions with G protein-coupled receptors. Recently many data have appeared about the role of this phospholipid in humans, but pathways of LPA biosynthesis and mechanisms of its action remain unclear. This review presents modern concepts about biosynthesis, reception, and biological activity of LPA in humans. Natural and synthetic LPA analogs are considered in the view of their possible use in pharmacology as agonists and/or antagonists of G protein-coupled receptors of LPA.
Subject(s)
Lysophospholipids/metabolism , Humans , Lipase/genetics , Lipase/metabolism , Lysophospholipids/biosynthesis , Lysophospholipids/physiology , Multienzyme Complexes/genetics , Multienzyme Complexes/metabolism , Phosphodiesterase I/genetics , Phosphodiesterase I/metabolism , Phosphoric Diester Hydrolases , Pyrophosphatases/genetics , Pyrophosphatases/metabolism , Receptors, G-Protein-Coupled/metabolismABSTRACT
The induction, regeneration, and biolistic sensitivities of different genotypes of common wheat (Triticum aestivum L.) have been determined in order to develop an efficient system for transformation of Russian cultivars of spring wheat. Short-term (two days) cold treatment (4 degrees C) has been demonstrated to distinctly increase the frequency of morphogenetic callus induction. The optimal phytohormonal composition of the nutrient medium ensuring an in vitro regeneration rate of the common wheat cultivar Lada as high as 90% has been determined. The optimal temporal parameters of genetic transformation of wheat plants (10-14 days of culturing after initiation of a morphogenetic callus) have been determined for two transformation methods: biolistic without precipitated DNA and transformation with the plasmid psGFP-BAR. Analysis of the transient expression of the gfp gene has confirmed that 14 days of culturing is the optimal duration.
Subject(s)
Plants, Genetically Modified/genetics , Transformation, Genetic , Triticum/genetics , Genotype , Plants, Genetically Modified/cytology , Plants, Genetically Modified/growth & development , Regeneration/genetics , Russia , Triticum/cytology , Triticum/growth & developmentABSTRACT
Expression of the desC gene coding for acyl-lipid delta(9) desaturase of thermophilic cyanobacterium Synechocystis sp. PCC6803 was studied in Escherichia coli cells. In a hybrid gene constructed (desC-licBM3), a sequence of the native acyl-lipid delta(9) desaturase was fused in frame with the reporter gene coding for thermostable lichenase. Lichenase contained in the hybrid protein simplified selection and analysis of the expression of membrane desaturase in the heterologous host. Comparisons of the expression for the native and hybrid genes in bacterial cells showed that lichenase remained active and thermostable in the hybrid protein, while desaturase retains the capability of introducing a double bound in the corresponding position of fatty acids.
