CLONING AND EXPRESSION ANALYSIS OF D-HORDEIN HYBRID PROMOTER ISOLATED FROM BARLEY (HORDEUM VULGARE L.)

Two variants of D-hordein promoter from barley ( Hordeum vulgare L.) were selected to generate hybrid promoter by using bioinformatics tools. Sequence analysis of both D-hordein promoter variants revealed several crucial cis regulatory elements and transcription factors binding sites within the promoter region. Various regulatory motifs related to endosperm specific expression were located in D-hordein promoter fragments. Both variants were separately cloned in TA vector ( pTZ57R/T) and then joined to get the complete hybrid promoter (D horH). Hybrid D-hordein promoter was further cloned in expression vector pGR1through adaptor ligation. Transient GUS assay revealed that hybrid promoter exhibited endosperm specific expression in wheat. From the study it is demonstrated that hybrid promoter ( D horH) may be used to derive endosperm specific expression in monocots. The present work provides an important insight in the designing of hybrid monocot promoters to improve multiple traits in crops without facing IPR issues.