Tandem repetition of baculovirus ie1 promoter results in upregulation of transcription

Kojima, Katsura; Hayakawa, Tohru; Asano, Shin Ichiro; Bando, Hideaki

doi:10.1007/s007050170101

Cited by 10 publications

(12 citation statements)

References 13 publications

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“…The Nanglai-BmNPV ie-1 ORF sequence was rich in AT, with 63.2% AT and 36.8% GC. The transcription start site (+1) 21 was located 50 bp upstream of the translation initiation codon, ATG. The termination codon, TAA, was located at the position +1802 to +1804 followed by a 3 untranslated region containing a putative polyadenylation site (AATAAA) at the position +2006 to +2011 (Fig.…”

Section: Resultsmentioning

confidence: 99%

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Chotimanothum¹,

Pinpart²,

Ngernsiri³

2011

ScienceAsia

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The immediate early 1 (ie-1) gene is essential for DNA replication of nucleopolyhedrosis virus. Here, the gene ie-1 from nucleopolyhedrovirus (Nanglai-BmNPV) infecting a local silkworm (Bombyx mori), Nanglai strain, was cloned and sequenced. The gene contained a 1755 bp open reading frame encoding a deduced protein of 584 amino acids, molecular weight 66.9 kDa, and isoelectric point 5.79. The nucleotide and amino acid sequences of Nanglai-BmNPV ie-1 gene showed, respectively, 98 and 97% identity to those of the BmNPV ie-1 genes. The identity of the Nanglai-BmNPV IE-1 deduced amino acid sequence with IE-1 protein sequences of other 37 known Lepidopteran NPVs varied from 20% up to 97%. The phylogenic tree established with the IE-1 protein sequences of the Lepidopteran NPVs showed similar topology to those previously reported using highly conserved genes. Interestingly, the tree obtained with IE-1 clearly shows that the Lepidopteran NPVs group II clade can be subdivided into three subgroups.

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Section: Resultsmentioning

confidence: 99%

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Chotimanothum¹,

Pinpart²,

Ngernsiri³

2011

ScienceAsia

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“…This is the first report to identify an ecdysone-responsive element in a baculoviral gene promoter. (Kojima et al, 2001) by digestion with Eco RI, blunt-ended, and treated with Sac I. The DNA fragments containing the ie1 promoter sequence were cloned between the Hin dIII / blunt and Sac I sites in PGV-p (Toyo Ink MFG.…”

Section: Resultsmentioning

confidence: 99%

“…BmN cells were transfected with plasmid DNA as described elsewhere (Kojima et al, 2001). After the transfection, the cells were maintained for 48 hrs in TC-100 medium containing alpha-ecdysone (Sigma) or 20-hydroxyecdysone (Sigma) at an implicit concentration as indicated in the figure legends.…”

Section: Transfection and Reporter Assaymentioning

confidence: 99%

“…After the transfection, the cells were maintained for 48 hrs in TC-100 medium containing alpha-ecdysone (Sigma) or 20-hydroxyecdysone (Sigma) at an implicit concentration as indicated in the figure legends. Then, luciferase activity in the cells was measured according to a method described elsewhere (Kojima et al, 2001). Ecdysteroids were dissolved in ethanol, then 0.1 mg/ml stock solutions were made in dH 2 O, and serially diluted with TC-100 culture medium to make the culture medium containing ecdysteroids at the implicit concentration shown in the figures.…”

Section: Transfection and Reporter Assaymentioning

confidence: 99%

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Ecdysone response element in a baculovirus immediate-early gene, ie1, promoter

et al. 2007

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A computer-assisted analysis identified tentative target sequences for regulatory proteins including ecdysone-inducible factors such as BmFTZ-F1 and Broad-Complex Z4 (BR-C Z4) in the ie1 promoter of BmNPV. A transient expression experiment using BmN cells and a series of truncated ie1 promoter constructs demonstrated that the activity of the ie1 promoter responded to alpha-ecdysone and 20-hydroxyecdysone, which required a tridecameric nucleotide stretch (ie1EcRE, 5'-GTGTTATCGACCT-3') homologous to the ecdysone response element reported for Drosophila (DmEcRE). RT-PCR demonstrated the expression of BmEcR and BmUSP, which are required as ecdysone-specific activators for EcRE-mediated activation, in BmN cells. Furthermore, the ie1EcRE-mediated response was confirmed by using a recombinant BmNPV possessing a luciferase gene under the control of the ie1 promoter with or without ie1EcRE. This is the first report of an ecdysone response element in a baculoviral gene promoter. These results also suggested that the regulation of the ie1 by ecdysone may militate viral replication at least under certain conditions during natural infections in vivo.

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“…Linkage of promoters of different nature in tandem has been tested in plants, insects and lactobacilli, and expression level was increased (Kay et al. 1987; Kojima et al. 2001; Wei et al.…”

Section: Introductionmentioning

confidence: 99%

Usage of an intronic promoter for stable gene expression in Saccharomyces cerevisiae

Shen

Jiang

et al. 2005

Lett Appl Microbiol

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Aims: To construct expression vectors capable of switching promoters under different metabolic circumstances to obtain stable gene expression. Methods and Results: In this study, we designed a series of constructs for the expression of the chicken lactate dehydrogenase (cldh) gene under the control of galactose-inducible GAL1 promoter and the high glucose-inducible HXT1 promoter in Saccharomyces cerevisiae. In one construct, the HXT1 promoter was placed between artificial splicing sequences to function as an intronic promoter. We checked all constructs for the usage of promoters by reverse transcriptional polymerase chain reaction and assayed the expression level of the reporter gene under different culturing conditions. In the presence of galactose, when the GAL1 promoter was linked with the intronic HXT1 promoter, the cldh gene showed 1AE5-fold activity compared with single GAL1 promoter, while in the presence of glucose, the construct showed over twofold activity compared with that without splicing sequences. Conclusion: The intronic HXT1 promoter could be induced by the presence of high glucose concentration. Significance and Impact of the Study: This is the first report detailing the use of an intronic promoter in the construction of stable expression vectors and the novel system could serve as a model of expression vectors for fermentation or other purposes.

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Tandem repetition of baculovirus ie1 promoter results in upregulation of transcription

Cited by 10 publications

References 13 publications

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Ecdysone response element in a baculovirus immediate-early gene, ie1, promoter

Usage of an intronic promoter for stable gene expression in Saccharomyces cerevisiae

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