Abstract
Although progression of genome-based techniques has been revam** several areas of genetic engineering, reliable and efficient procedures are expected to unveil structural and functional information of genes. Many methods such as chromosome walking and molecular cloning that are used to recognize unknown flanking sequences are effortful and time-consuming. Here, we report the identification of an unknown upstream regulatory region of actin gene from Plectranthus amboinicus and eight other medicinal plants using thermal asymmetric interlaced PCR (TAIL PCR). As actin is a ubiquitous protein that plays a significant role in developmental stages of plants, we set out to isolate the 5′ flanking region of the actin gene. Three heterologous gene-specific primers were designed based on plant Arabidopsis actin conserved sites, and arbitrary degenerate primers were used for the isolation of putative promoter sequence. Successful amplification was observed in most of the plants tested, thus proving that TAIL PCR is an efficient, effective, and economic procedure for the isolation of promoter sequences from various plants.
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The data and the materials used in the current study are available with the corresponding author and can be produced on request.
Abbreviations
- TAIL PCR:
-
Thermal asymmetric interlaced PCR
- AD primers:
-
Arbitrary degenerate primers
- GSP:
-
Gene-specific primers
- µM:
-
Micromolar
- OD:
-
Optical density
- ARE:
-
Anaerobic responsive element
- GARE:
-
Gibberellin-responsive element
- I-Box:
-
Light responsive element
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Acknowledgements
We are immensely grateful to the management of Sri Ramachandra Institute of Higher Education and Research (DU) for Founder Chancellor Shri NPV Ramasamy Udayar Fellowship in support of the study and providing necessary infrastructure.
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ECSM preformed laboratory work and manuscript writing. AV helped in conception and designed the study, analysed the data, and manuscript preparation.
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Accession numbers: GenBank accession numbers for the nucleotide sequence ptcoac.AD4 from Plectranthus amboinicus are OM892829. GenBank accession numbers for the nucleotide sequence CA639_SP from Senna auriculata are ON000299. GenBank accession numbers for the nucleotide sequence CS639_SP from Coriandrum sativum are ON000300.
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Evangelene Christy, S.M., Arun, V. Isolation of actin regulatory region from medicinal plants by thermal asymmetric interlaced PCR (TAIL PCR) and its bioinformatic analysis. Braz. J. Bot 47, 67–78 (2024). https://doi.org/10.1007/s40415-023-00971-z
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DOI: https://doi.org/10.1007/s40415-023-00971-z