Structure of Alpha-Gliadin Multigene and Construction of Efficient Hairpin RNAi Molecule against Gluten Enteropathy

Abstract

Celiac disease is known as gluten enteropathy, caused by damaging of small intestinal epithelium cells following gluten consumption. Gluten is classified in two families of glutenin and gliadin. Gliadin is divided into α , γ and ω groups. One of the most effective methods to rectify or minimize gene expression is RNAi technology. The present study has been conducted to the structure of α -gliadin gene to produce proprietary RNAi cassette for silencing wheat α -gliadin gene. So nucleotide sequence of α -gliadin gene involved in design for PCR amplification by online primer-blast software. The amplification was extracted from agarose gel and ligated to pTG19 cloning vector. After cloning of the recombinant plasmid in E. coli , they were sequenced. Then they were used for construction of specific and efficient RNAi cassette. The produced vector by this strategy is considered as an effective step in developing genetically engineered gluten-less or low content gluten wheat.