Isolation and sequence analysis of napin seed specific promoter from Iranian Rapeseed (Brassica napus L.)

Rapeseed (Brassica napus L.) has become an important crop during the last 30years. In addition to a high lipid level, the seeds also have a significant protein content, which constitutes 20–25% of the dry seed weight. The synthesis of storage proteins is primarily controlled at transcriptional level...

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Bibliographic Details
Published in:Gene 2015-06, Vol.563 (2), p.160-164
Main Authors: Sohrabi, Maryam, Zebarjadi, Alireza, Najaphy, Abdollah, Kahrizi, Danial
Format: Article
Language:English
Subjects:
Base Sequence
Brassica napus
Brassica napus - genetics
Cloning
Molecular Sequence Data
Napin promoter
Phylogeny
Plant Proteins - genetics
Promoter Regions, Genetic
Seed-specific promoter
Seeds - genetics
Sequence Analysis
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Summary:Rapeseed (Brassica napus L.) has become an important crop during the last 30years. In addition to a high lipid level, the seeds also have a significant protein content, which constitutes 20–25% of the dry seed weight. The synthesis of storage proteins is primarily controlled at transcriptional level and seed-specific expression has been shown to be conferred upon the promoter regions of many storage protein genes. Napin is one of the main storage proteins in rapeseed's embryo that is produced in seed developing stage. Its promoter region located at 5′ upstream of the napin gene has already been isolated (GenBank number, EU416279.1). In current research, seed-specific promoter (napin) of Iranian B. napus L. was isolated from the genomic DNA and cloned into pBI121 plant binary vector to use in future researches. For this purpose, the napin promoter was amplified by PCR method using specific primers, cloned in pSK+ vector and sequenced. Sequencing analysis showed that the cloned promoter contained all of conserved motifs such as TATA box (TATAAA), RY repeats (CATGCA), dist-B (TCAAACACC) and prox-B elements (GCCACTTGTC), G-box (CACGTG) and CAAT Motifs, which constituted the seed-specific promoter activity and according to this analysis, the seed-specific promoter activity of cloned sequence was predicted. Based on sequence distances of nucleotide sequences, our sequence had the highest similarity (99.8%) whit B. napus sequence (with EU416279.1 accession number). Finally the promoter obtained might be interesting not only as a useful tool for biotechnological application but also for fundamental research. •We isolated Napin promoter from an Iranian variety of B. napus for the first time.•We found some motifs that they had key roles in seed-specific activity.•Isolated promoter had differences with other napin promoters in B. napus.•The promoter had differences with napin promoter of B. juncea, oleracea and rapa.
ISSN:0378-1119
1879-0038
DOI:10.1016/j.gene.2015.03.040