Analysis of promoter activity in transgenic plants by normalizing expression with a reference gene: anomalies due to the influence of the test promoter on the reference promoter

2009 ◽  
Vol 34 (6) ◽  
pp. 953-962 ◽  
Author(s):  
Simran Bhullar ◽  
Suma Chakravarthy ◽  
Deepak Pental ◽  
Pradeep Kumar Burma
Keyword(s):  
Transgenic Plants ◽  
Reference Gene ◽  
Promoter Activity

scholarly journals High-Level and Ubiquitous Expression of the Rice Cytochromec Gene OsCc1 and Its Promoter Activity in Transgenic Plants Provides a Useful Promoter for Transgenesis of Monocots

2002 ◽  
Vol 129 (4) ◽  
pp. 1473-1481 ◽  
Author(s):  
In-Cheol Jang ◽  
Won-Bin Choi ◽  
Kyung-Hee Lee ◽  
Sang Ik Song ◽  
Baek Hie Nahm ◽  
...  
Keyword(s):  
Transgenic Plants ◽  
Promoter Activity ◽  
High Level

scholarly journals Long terminal repeats (LTR) and transcription factors regulate PHRE1 and PHRE2 activity in Moso bamboo under heat stress

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Pradeep K. Papolu ◽  
Muthusamy Ramakrishnan ◽  
Qiang Wei ◽  
Kunnummal Kurungara Vinod ◽  
Long-Hai Zou ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Heat Stress ◽  
Transgenic Plants ◽  
Promoter Activity ◽  
Moso Bamboo ◽  
Ltr Retrotransposons ◽  
In Planta ◽  
Environmental Stress Response ◽  
Long Terminal Repeats ◽  
Camv35s Promoter

Abstract Background LTR retrotransposons play a significant role in plant growth, genome evolution, and environmental stress response, but their regulatory response to heat stress remains unclear. We have investigated the activities of two LTR retrotransposons, PHRE1 and PHRE2, of moso bamboo (Phyllostachys edulis) in response to heat stress. Results The differential overexpression of PHRE1 and PHRE2 with or without CaMV35s promoter showed enhanced expression under heat stress in transgenic plants. The transcriptional activity studies showed an increase in transposition activity and copy number among moso bamboo wild type and Arabidopsis transgenic plants under heat stress. Comparison of promoter activity in transgenic plants indicated that 5’LTR promoter activity was higher than CaMV35s promoter. Additionally, yeast one-hybrid (Y1H) system and in planta biomolecular fluorescence complementation (BiFC) assay revealed interactions of heat-dependent transcription factors (TFs) with 5’LTR sequence and direct interactions of TFs with pol and gag. Conclusions Our results conclude that the 5’LTR acts as a promoter and could regulate the LTR retrotransposons in moso bamboo under heat stress.

scholarly journals Expression analysis of acdS-gene of Pseudomonas putida B-37 in transgenic plants Nicotiana tabacum

2019 ◽  
pp. 45-53 ◽  
Author(s):  
Alesia A. Melnikava ◽  
Alena A. Khramtsova ◽  
Katsiaryna S. Karaleva ◽  
Daria A. Rutkevich ◽  
Tatsiana A. Kukulianskaya
Keyword(s):  
Transgenic Plants ◽  
Reference Gene ◽  
Target Gene ◽  
Specific Activity ◽  
Acc Deaminase ◽  
Pcr Analysis ◽  
Acds Gene ◽  
The Difference ◽  
High Level ◽  
Present Gene

In current work was realized the transfer of recombinant plasmid pBI121acdS, carring P. putida B-37 bacterial acdSgene to the A. tumefaciens AGL0 cells. Transgenic plants of N. tabacum were created by agrobacterial transformation. Integration of P. putida B-37 bacterial acdS-gene to transgenic plants of N. tabacum was verified by PCR analysis, using specific primers to present gene. Presence of target acdS-gene in transgenic plants genome was proved by RT-PCR analysis. With help of  Real-time PCR was shown the difference between reference gene and target P. putida B-37 bacterial acdSgene expression. Expression of target gene exceeded reference gene in 1.27 times, those fact proved expression of acdS-gene in plants on high level. Expression of the heterologous gene in N. tabacum plants was also proved by biochemical method of ACC-deaminase specific activity define.

Promoter deletion analysis reveals root-specific expression of the alkenal reductase gene (OsAER1) in Oryza sativa

2019 ◽  
Vol 46 (4) ◽  
pp. 376
Author(s):  
Aniversari Apriana ◽  
Atmitri Sisharmini ◽  
Hajrial Aswidinnoor ◽  
Kurniawan R. Trijatmiko ◽  
Sudarsono Sudarsono
Keyword(s):  
Oryza Sativa ◽  
Transgenic Plants ◽  
Promoter Activity ◽  
Deletion Analysis ◽  
Specific Expression ◽  
Cis Acting ◽  
Promoter Fragment ◽  
Gus Reporter ◽  
Promoter Deletion Analysis

Root-specific promoters are useful in plant genetic engineering, primarily to improve water and nutrient absorption. The aim of this study was to clone and characterise the promoter of the Oryza sativa L. alkenal reductase (OsAER1) gene encoding 2-alkenal reductase, an NADPH-dependent oxidoreductase. Expression analysis using quantitative real-time PCR confirmed the root-specific expression of the OsAER1 gene. Subsequently, a 3082-bp fragment of the OsAER1 promoter was isolated from a local Indonesian rice cultivar, Awan Kuning. Sequencing and further nucleotide sequence analysis of the 3082-bp promoter fragment (PA-5) revealed the presence of at least 10 root-specific cis-regulatory elements putatively responsible for OsAER1 root-specific expression. Using the 3082-bp promoter fragment to drive the expression of the GUS reporter transgene confirmed that the OsAER1 promoter is root-specific. Further, the analysis indicated that OsAER1 promoter activity was absent in leaves, petioles and shoots during sprouting, vegetative, booting and generative stages of rice development. In contrast, the promoter activity was present in anthers and aleurone layers of immature seeds 7–20 days after anthesis. Moreover, there was no promoter activity observed in the aleurone layers of mature seeds. The OsAER1 promoter activity is induced by Al-toxicity, NaCl and submergence stresses, indicating the OsAER1 promoter activity is induced by those stresses. Exogenous treatments of transgenic plants carrying the PA-5 promoter construct with abscisic acid and indoleacetic acid also induced expression of the GUS reporter transgene, indicating the role of plant growth regulators in controlling OsAER1 promoter activity. Promoter deletion analysis was conducted to identify the cis-acting elements of the promoter responsible for controlling root-specific expression. The GUS reporter gene was fused with various deletion fragments of the OsAER1 promoter and the resulting constructs were transformed in rice plants to generate transgenic plants. The results of this analysis indicated that cis-acting elements controlling root-specific expression are located between −1562 to −1026bp of the OsAER1 CDS. Here we discusses the results of the conducted analyses, the possible role of OsAER1 in rice growth and development, possible contributions and the potential usage of these findings in future plant research.

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