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Molecular Cloning and Expression Analysis of MYB-Related Transcription Factor Gene, ScMYB76 from Sugarcane (Saccharum Hybrid)


Affiliations
1 Department of Biotechnology, Karpagam University (Karpagam Academy of Higher Education), Eachanari, Coimbatore - 641021, Tamil Nadu, India
 

Background/Objectives: In grass family, sugarcane has been gaining importance due to its role in sugar and biofuel production. We previously reported MYB Transcription Factor (TF) family of genes that regulates almost all biological, physical and biochemical functions in sugarcane. Thus, the present study aimed to characterize an MYB-related Transcription Factor (TF) gene, of ScMYB76 from sugarcane and its expression during water-deficit and salt stress induced condition. Methods: ScMYB76 gene specific primers were designed and used to isolate by PCR from sugarcane leaf tissue. The isolated gene was cloned and sequenced for sequence-structure analysis using various computational tools. To analyze the sequence-to-structure-to-function paradigm, comparative modeling, docking and validation were performed using I-TASSER, TFmodeller and 3D footprint platforms, respectively. The expression pattern during the induced stress condition was analyzed by quantitative RT-PCR. Findings: A cDNA encloding a putative MYB-related TF, ScMYB76 was cloned from sugarcane leaves and consisted of 636 bp Open Reading Frame (ORF) which encodes 212 amino acid length protein with the calculated molecular mass of 23.49 KDa and a pI of 10.2. The deduced ScMYB76 protein was predicted to consist of a single helix-turn-helix MYB-like motif and other residues which are highly conserved among other MYB-related TFs. Comparative modeling and docking of ScMYB76 with MYBCORE cis-motif was generated and evaluated by structure quality assessment parameters. Analysis of expression pattern of ScMYB76 indicates an important function of this gene during abiotic stress in sugarcane. Applications/Improvements: The isolated and analyzed TF gene, ScMYB76 from sugarcane will add a valuable gene resource for crop improvement in regard to abiotic stress.

Keywords

Docking, Expression, MYB-Related, Stress, Sugarcane, Transcription Factor.
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  • Molecular Cloning and Expression Analysis of MYB-Related Transcription Factor Gene, ScMYB76 from Sugarcane (Saccharum Hybrid)

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Authors

Hemalatha Balakrishnan
Department of Biotechnology, Karpagam University (Karpagam Academy of Higher Education), Eachanari, Coimbatore - 641021, Tamil Nadu, India
Prabu Gajjeraman
Department of Biotechnology, Karpagam University (Karpagam Academy of Higher Education), Eachanari, Coimbatore - 641021, Tamil Nadu, India
Yasir Umar Pattiwala
Department of Biotechnology, Karpagam University (Karpagam Academy of Higher Education), Eachanari, Coimbatore - 641021, Tamil Nadu, India

Abstract


Background/Objectives: In grass family, sugarcane has been gaining importance due to its role in sugar and biofuel production. We previously reported MYB Transcription Factor (TF) family of genes that regulates almost all biological, physical and biochemical functions in sugarcane. Thus, the present study aimed to characterize an MYB-related Transcription Factor (TF) gene, of ScMYB76 from sugarcane and its expression during water-deficit and salt stress induced condition. Methods: ScMYB76 gene specific primers were designed and used to isolate by PCR from sugarcane leaf tissue. The isolated gene was cloned and sequenced for sequence-structure analysis using various computational tools. To analyze the sequence-to-structure-to-function paradigm, comparative modeling, docking and validation were performed using I-TASSER, TFmodeller and 3D footprint platforms, respectively. The expression pattern during the induced stress condition was analyzed by quantitative RT-PCR. Findings: A cDNA encloding a putative MYB-related TF, ScMYB76 was cloned from sugarcane leaves and consisted of 636 bp Open Reading Frame (ORF) which encodes 212 amino acid length protein with the calculated molecular mass of 23.49 KDa and a pI of 10.2. The deduced ScMYB76 protein was predicted to consist of a single helix-turn-helix MYB-like motif and other residues which are highly conserved among other MYB-related TFs. Comparative modeling and docking of ScMYB76 with MYBCORE cis-motif was generated and evaluated by structure quality assessment parameters. Analysis of expression pattern of ScMYB76 indicates an important function of this gene during abiotic stress in sugarcane. Applications/Improvements: The isolated and analyzed TF gene, ScMYB76 from sugarcane will add a valuable gene resource for crop improvement in regard to abiotic stress.

Keywords


Docking, Expression, MYB-Related, Stress, Sugarcane, Transcription Factor.



DOI: https://doi.org/10.17485/ijst%2F2016%2Fv9i37%2F126842