A polyclonal antibody against a recombinantly expressed Triticum aestivum RHT-D1A protein

作者：Izat Smekenov, Sanzhar Alybayev, Temurkhan Ayupov, Guliza Rakhmatullaeva, Amangeldy Bissenbaev · 发表于：Journal of Genetic Engineering and Biotechnology · 年份：2020 · DOI：10.1186/s43141-020-00072-4 · 被引用次数：4 · 研究领域：Plant Molecular Biology Research、Wheat and Barley Genetics and Pathology、Plant nutrient uptake and metabolism

BACKGROUND: Reduced height-1 dwarfing alleles affect DELLA proteins belonging to a family of putative transcriptional regulators that modulate plant growth and development. The Arabidopsis thaliana genome encodes five DELLA proteins, whereas monocot plants, such as rice, barley, and wheat, each have a single DELLA protein. In wheat, wild-type Rht-B1a and Rht-D1a genes encode DELLA proteins and have many alleles that contain lesions. Among them, Rht-B1b and Rht-D1b are the most common mutant dwarfing alleles, which have played a key part in the creation of high-yielding wheat varieties. Despite their fundamental roles in plant biology, until now, DELLA proteins in wheat have been mainly researched regarding the phenotypic effect of defective Rht mutants on yield-related traits, without studies on the underlying mechanisms. The RHT-1 protein has yet to be detected in wheat tissues, owing to a lack of appropriate molecular tools for characterization of RHT function and protein interactions in signal transduction. This study is focused on the production of a polyclonal antibody to the wheat RHT-D1A protein. RESULTS: To generate the anti-RHT-D1A antibody, we expressed and purified soluble 6xHis-tagged RHT-D1A. The purified recombinant RHT-D1A was injected into New Zealand white rabbits to generate polyclonal antiserum. The polyclonal anti-RHT-D1A antibody was purified by ammonium sulfate precipitation, followed by affinity chromatography on protein A-agarose beads. The purified po...