Dataset Information

Novel strong promoter of antimicrobial peptides gene pro-SmAMP2 from chickweed (Stellaria media).

ABSTRACT: In a previous study we found that in chickweed the expression level of the pro-SmAMP2 gene was comparable or even higher to that of the ?-actin gene. This high level of the gene expression has attracted our attention as an opportunity for the identification of novel strong promoters of plant origin, which could find its application in plant biotechnology. Therefore, in the present study we focused on the nucleotide sequence identification and the functional characteristics of the pro-SmAMP2 promoter in transgenic plants.In chickweed (Stellaria media), a 2120 bp promoter region of the pro-SmAMP2 gene encoding antifungal peptides was sequenced. Six 5'-deletion variants -2120, -1504, -1149, -822, -455, and -290 bp of pro-SmAMP2 gene promoter were fused with the coding region of the reporter gene gusA in the plant expression vector pCambia1381Z. Independent transgenic plants of tobacco Nicotiana tabacum were obtained with each genetic structure. GUS protein activity assay in extracts from transgenic plants showed that all deletion variants of the promoter, except -290 bp, expressed the gusA gene. In most transgenic plants, the GUS activity level was comparable or higher than in plants with the viral promoter CaMV 35S. GUS activity remains high in progenies and its level correlates positively with the amount of gusA gene mRNA in T3 homozygous plants. The activity of the ?ro-SmAMP2 promoter was detected in all organs of the transgenic plants studied, during meiosis and in pollen as well.Our results show that the ?ro-SmAMP2 promoter can be used for target genes expression control in transgenic plants.

SUBMITTER: Komakhin RA

PROVIDER: S-EPMC4870781 | biostudies-literature | 2016 May

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Novel strong promoter of antimicrobial peptides gene pro-SmAMP2 from chickweed (Stellaria media).

Komakhin Roman A RA Vysotskii Denis A DA Shukurov Rahim R RR Voblikova Vera D VD Komakhina Vera V VV Strelnikova Svetlana R SR Vetchinkina Ekaterina M EM Babakov Alexey V AV

BMC biotechnology 20160518 1

<h4>Background</h4>In a previous study we found that in chickweed the expression level of the pro-SmAMP2 gene was comparable or even higher to that of the β-actin gene. This high level of the gene expression has attracted our attention as an opportunity for the identification of novel strong promoters of plant origin, which could find its application in plant biotechnology. Therefore, in the present study we focused on the nucleotide sequence identification and the functional characteristics of ...[more]

PMID: 27189173

Similar Datasets

Project description:BackgroundTissue specific promoters may be utilized for a variety of applications, including programmed gene expression in cell types, tissues and organs of interest, for developing different cell culture models or for use in gene therapy. We report a novel, tissue-specific promoter that was identified and engineered from the native upstream regulatory region of the human gene NDUFV1 containing an endogenous retroviral sequence.ResultsAmong seven established human cell lines and five primary cultures, this modified NDUFV1 upstream sequence (mNUS) was active only in human undifferentiated germ-derived cells (lines Tera-1 and EP2102), where it demonstrated high promoter activity (approximately twice greater than that of the SV40 early promoter, and comparable to the routinely used cytomegaloviral promoter). To investigate the potential applicability of the mNUS promoter for biotechnological needs, a construct carrying a recombinant cytosine deaminase (RCD) suicide gene under the control of mNUS was tested in cell lines of different tissue origin. High cytotoxic effect of RCD with a cell-death rate approximately 60% was observed only in germ-derived cells (Tera-1), whereas no effect was seen in a somatic, kidney-derived control cell line (HEK293). In further experiments, we tested mNUS-driven expression of a hyperactive Sleeping Beauty transposase (SB100X). The mNUS-SB100X construct mediated stable transgene insertions exclusively in germ-derived cells, thereby providing further evidence of tissue-specificity of the mNUS promoter.ConclusionsWe conclude that mNUS may be used as an efficient promoter for tissue-specific gene expression in human germ-derived cells in many applications. Our data also suggest that the 91 bp-long sequence located exactly upstream NDUFV1 transcriptional start site plays a crucial role in the activity of this gene promoter in vitro in the majority of tested cell types (10/12), and an important role--in the rest two cell lines.

Dataset Information

Novel strong promoter of antimicrobial peptides gene pro-SmAMP2 from chickweed (Stellaria media).

Publications

Novel strong promoter of antimicrobial peptides gene pro-SmAMP2 from chickweed (Stellaria media).

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets