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Molecular cloning and expression analysis of the transcription factor CobHLH40 from Cornus officinalis on the basis of de novo transcriptome sequencing

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Abstract

Cornus officinalis is a traditional Chinese herbal medicine rich in active substances, such as iridoid glycosides, esters, polysaccharides, and tannins. C. officinalis replenishes the liver and kidney and retracts and removes phlegm. In this study, the candidate transcription factor bHLH40 from C. officinalis (CobHLH40) was predicted to be involved in iridoid glycoside biosynthesis. CobHLH40 was cloned from C. officinalis. Bioinformatics analysis was performed using the Protparatam, the ProtScale, and the SOPMA. CobHLH40 had a cDNA length of 2100 bp and a relative molecular mass of 63.32 kDa, and encoded 573 amino acids. The subcellular localization analysis indicated that CobHLH40 may be located in the nucleus. The comparison of multiple sequence alignments and phylogenetic relationships showed that the CobHLH40 was similar to the MYC2 of other plants, and that CobHLH40 had high homology with the MYC2 of Sesamum indicum. A prokaryotic expression vector was successfully constructed and transformed into the DE3 cells. The induction and expression of the fusion protein of CobHLH40 need further confirmation. The cloning and bioinformatics analysis results of CobHLH40, which may be related to iridoid glycoside synthesis, were screened and predicted for the first time on the basis of C. officinalis transcriptome sequencing. The temporal and spatial expression patterns of C. officinalis were also established. The CobHLH40 prokaryotic expression system provides a basis to further study its function.

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Abbreviations

NR:

Non-redundant

RT-PCR:

Reverse transcription polymerase chain reaction

bHLH:

Basic helix-loop-helix

GO:

Gene Ontology

qRT-PCR:

Quantitative real-time PCR

IPTG:

Isopropyl-d-1-thiogalactopyranoside

KEGG:

Kyoto Encyclopedia of Genes and Genomes

LB medium:

Luria–Bertani medium

MYC:

Myelocytomatosis proteins

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (U1404829) and the key project at central government level: The ability establishment of sustainable use for valuable Chinese medicine resources (2060302). This work was also supported by the special fund for the construction of technical system of traditional Chinese medicine industry in Henan province.

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Author notes

  1. Dianyun Hou and Wenbin Wang have contributed equally to this work.

Authors and Affiliations

  1. Agricultural College, Henan University of Science and Technology, Luoyang, 471023, People’s Republic of China

    Dianyun Hou, Mengmeng Yang, Zhihong Li, Jingya Xu, Long Wang, Zhenjie Fan & Yaoyao Wang

  2. The Luoyang Engineering Research Center of Breeding and Utilization of Dao-Di Herbs, Luoyang, 471023, People’s Republic of China

    Dianyun Hou, Mengmeng Yang, Zhihong Li, Jingya Xu, Long Wang, Zhenjie Fan & Yaoyao Wang

  3. College of Life Sciences, Shanxi Agricultural University, Taigu, 030801, People’s Republic of China

    Wenbin Wang

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  1. Dianyun Hou
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Hou, D., Wang, W., Yang, M. et al. Molecular cloning and expression analysis of the transcription factor CobHLH40 from Cornus officinalis on the basis of de novo transcriptome sequencing. Plant Biotechnol Rep 14, 419–428 (2020). https://doi.org/10.1007/s11816-020-00623-8

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