Isolating Barley (<i>Hordeum vulgare</i> L.) B1 Hordein Gene Promoter and Using Sequencing Analaysis For The Identification of Conserved Regulatory Elements By Bioinformatic Tools

Abstract

Gene expression is a complex multi-step process. For the efficient expression of foreign genes in plants, it is essential to optimize every step of the process for the plant machinery, which includes choosing suitable promoters. Promoters play the most important role in determining the temporal and spatial expression pattern and transcript level of a gene. Some strong constitutive promoters, such as cauliflower mosaic virus 35s promoter, are widely used in plant genetic engineering research. However, the expression levels of the foreign genes in all tissues are often unsatisfied, but some of such problems can be may solved by using a strong seed-specific promoter to restrict gene expression to the seed only. The aim of this article was to characterize a B1 hordein-specific promoter. The promoter region of B1 hordein gene was isolated from the genomic DNA of Walfajre and Alger barley by polymerase chain reaction. Sequence analysis showed that the cloned fragment B hordein promoter (BHP) - contained motifs like TATA box, (CA)n box, ACGT motif, AAAG motif, GCN4-like motif and Ebox, which constituted the seed-specific promoter activity. It was therefore concluded that B1 hordein promoters can be used to engineer and subsequently study stable seed specific gene expression in barley, and potentially to modify barley seeds through genetic engineering.

Keywords: Barley, Seed Specific Promoter, Motif, Hordein

African Journal of Biotechnology Vol. 11(29), pp. 7378-7387, 10 April, 2012