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Identification and use of the sugarcane bacilliform virus enhancer in transgenic maize.

Davies JP, Reddy V, Liu XL, Reddy AS, Ainley WM, Thompson M, Sastry-Dent L, Cao Z, Connell J, Gonzalez DO, Wagner DR - BMC Plant Biol. (2014)

Bottom Line: When the enhancer array was transformed into maize plants it caused an increase in accumulation of transcripts of genes near the site of integration in the genome.The SCBV-IM enhancer can activate transcription upstream or downstream of genes and in either orientation.It may be a useful tool to activate enhance from specific promoters or in activation tagging.

View Article: PubMed Central - PubMed

ABSTRACT

Background: Transcriptional enhancers are able to increase transcription from heterologous promoters when placed upstream, downstream and in either orientation, relative to the promoter. Transcriptional enhancers have been used to enhance expression of specific promoters in transgenic plants and in activation tagging studies to help elucidate gene function.

Results: A transcriptional enhancer from the Sugarcane Bacilliform Virus - Ireng Maleng isolate (SCBV-IM) that can cause increased transcription when integrated into the the genome near maize genes has been identified. In transgenic maize, the SCBV-IM promoter was shown to be comparable in strength to the maize ubiquitin 1 promoter in young leaf and root tissues. The promoter was dissected to identify sequences that confer high activity in transient assays. Enhancer sequences were identified and shown to increase the activity of a heterologous truncated promoter. These enhancer sequences were shown to be more active when arrayed in 4 copy arrays than in 1 or 2 copy arrays. When the enhancer array was transformed into maize plants it caused an increase in accumulation of transcripts of genes near the site of integration in the genome.

Conclusions: The SCBV-IM enhancer can activate transcription upstream or downstream of genes and in either orientation. It may be a useful tool to activate enhance from specific promoters or in activation tagging.

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Related in: MedlinePlus

SCBV promoter sequence (A) and fragments used in experiments (B). (A) The transcription start site was mapped by 5′ RACE and sequences in the 5′ untranslated region of the transcript are underlined (position 1 – 69). The putative TATA box is underlined at position −28 – -33. The SCBV-IM enhancer sequences are in bold. (B) The fragments of the SCBV-IM promoter used in transient assays are displayed. The position of the transcription start site (T) is shown.
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Fig2: SCBV promoter sequence (A) and fragments used in experiments (B). (A) The transcription start site was mapped by 5′ RACE and sequences in the 5′ untranslated region of the transcript are underlined (position 1 – 69). The putative TATA box is underlined at position −28 – -33. The SCBV-IM enhancer sequences are in bold. (B) The fragments of the SCBV-IM promoter used in transient assays are displayed. The position of the transcription start site (T) is shown.

Mentions: The activity of the SCBV-IM enhancer was demonstrated in transient assays first by identifying sequences that are necessary for high levels of transcription and then by identifying sequences that can enhance transcription from a heterologous promoter. The sequence of the SCBV-IM promoter is shown in Figure 2; the transcription start site was mapped by 5′ RACE.Figure 2


Identification and use of the sugarcane bacilliform virus enhancer in transgenic maize.

Davies JP, Reddy V, Liu XL, Reddy AS, Ainley WM, Thompson M, Sastry-Dent L, Cao Z, Connell J, Gonzalez DO, Wagner DR - BMC Plant Biol. (2014)

SCBV promoter sequence (A) and fragments used in experiments (B). (A) The transcription start site was mapped by 5′ RACE and sequences in the 5′ untranslated region of the transcript are underlined (position 1 – 69). The putative TATA box is underlined at position −28 – -33. The SCBV-IM enhancer sequences are in bold. (B) The fragments of the SCBV-IM promoter used in transient assays are displayed. The position of the transcription start site (T) is shown.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4302606&req=5

Fig2: SCBV promoter sequence (A) and fragments used in experiments (B). (A) The transcription start site was mapped by 5′ RACE and sequences in the 5′ untranslated region of the transcript are underlined (position 1 – 69). The putative TATA box is underlined at position −28 – -33. The SCBV-IM enhancer sequences are in bold. (B) The fragments of the SCBV-IM promoter used in transient assays are displayed. The position of the transcription start site (T) is shown.
Mentions: The activity of the SCBV-IM enhancer was demonstrated in transient assays first by identifying sequences that are necessary for high levels of transcription and then by identifying sequences that can enhance transcription from a heterologous promoter. The sequence of the SCBV-IM promoter is shown in Figure 2; the transcription start site was mapped by 5′ RACE.Figure 2

Bottom Line: When the enhancer array was transformed into maize plants it caused an increase in accumulation of transcripts of genes near the site of integration in the genome.The SCBV-IM enhancer can activate transcription upstream or downstream of genes and in either orientation.It may be a useful tool to activate enhance from specific promoters or in activation tagging.

View Article: PubMed Central - PubMed

ABSTRACT

Background: Transcriptional enhancers are able to increase transcription from heterologous promoters when placed upstream, downstream and in either orientation, relative to the promoter. Transcriptional enhancers have been used to enhance expression of specific promoters in transgenic plants and in activation tagging studies to help elucidate gene function.

Results: A transcriptional enhancer from the Sugarcane Bacilliform Virus - Ireng Maleng isolate (SCBV-IM) that can cause increased transcription when integrated into the the genome near maize genes has been identified. In transgenic maize, the SCBV-IM promoter was shown to be comparable in strength to the maize ubiquitin 1 promoter in young leaf and root tissues. The promoter was dissected to identify sequences that confer high activity in transient assays. Enhancer sequences were identified and shown to increase the activity of a heterologous truncated promoter. These enhancer sequences were shown to be more active when arrayed in 4 copy arrays than in 1 or 2 copy arrays. When the enhancer array was transformed into maize plants it caused an increase in accumulation of transcripts of genes near the site of integration in the genome.

Conclusions: The SCBV-IM enhancer can activate transcription upstream or downstream of genes and in either orientation. It may be a useful tool to activate enhance from specific promoters or in activation tagging.

Show MeSH
Related in: MedlinePlus