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Cloning, characterization, and transcriptional activity of β-actin promoter of African catfish (Clarias gariepinus)

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Abstract

Selection of suitable promoters is crucial for the efficient expression of exogenous genes in transgenic animals. Although one of the most effective promoters, the β-actin promoter, has been widely studied in fish species, it still remains unknown in the economical important African catfish (Clarias gariepinus). In this study, the β-actin promoter of African catfish (cgβ-actinP) was cloned and characterized. In addition, recombinant plasmid pcgβ-actinP-EGFP with enhanced green fluorescent protein (GFP) gene as the reporter gene was constructed to verify the transcriptional activity. We obtained a cgβ-actinP fragment length of 1405 bp, consisting 104 bp of the 5′ proximal promoter, 96 bp of the first exon, and 1205 bp of the first intron. Similar to those of other fish species, cgβ-actinP contains three key transcription regulatory elements (CAAT box, CArG motif, and TATA box). GFP-specific fluorescent signals were detected in chicken embryonic fibroblasts cells (DF-1 cells) transfected with pcgβ-actinP-EGFP, which was approximately 1.11 times of the positive control. In addition, GFP was effectively expressed in zebrafish larvae microinjected with linearized cgβ-actinP-EGFP, with expression rate reaching approximately 49.84%. Our data indicate that cgβ-actinP could be a potential candidate promoter in the practice of constructing “all fish” transgenic fish.

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The data sets supporting the results of this article are included within the article. Additional data and/or material related to this paper may be requested from the authors.

Abbreviations

cgβ-actinP :

African catfish (Clarias gariepinus) β-actin promoter

GFP:

Green fluorescent protein

FAO:

Food and agriculture organization of the United Nations

CDS:

Coding sequences

ZFIN:

Zebrafish information network

WT:

Wild type

DF-1 cells:

Chicken embryonic fibroblasts cells

CMV:

Cytomegalovirus

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Funding

This work was financial supported by National Key R&D Program of China (No. 2018YFD0900603), the Open Project of Key Laboratory of Environmental Biotechnology, CAS (kf2018005), and the Science and Technology Project of Wenzhou (2019ZX002-02).

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Author notes

  1. Maocang Yan and Guangxu Liu contribute equally to this manuscript.

Authors and Affiliations

  1. College of Animal Sciences, Zhejiang University, Hangzhou, 310058, PR China

    Sanghyok Ri, Wenho Hwang, Wei Shi, Yu Han, Yu Tang & Guangxu Liu

  2. College of Life Science, Kim Hyong Jik University of Education, Pyongyang, 99903, Democratic People’s Republic of Korea

    Sanghyok Ri

  3. Faculty of Chemistry, Kim II Sung University, Pyongyang, 99903, Democratic People’s Republic of Korea

    Sangryong Ri

  4. Zhejiang Mariculture Research Institute, Wenzhou, 325005, PR China

    Lining Zhang & Maocang Yan

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  1. Sanghyok Ri
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Contributions

SR: Conceptualization, investigation, methodology, writing-original draft. WH: Data curation, investigation. SR: Data curation, investigation. WS: Data curation. YH: Formal analysis, investigation. YT: Formal analysis, investigation. LZ: Formal analysis, investigation. MY: Conceptualization, project administration, supervision, funding acquisition. GL: Conceptualization, project administration, supervision, funding acquisition.

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Correspondence to Guangxu Liu.

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Ethics code permit No. ZJU2015-516-15, issued by the Animal Ethics Committee in the School of Medicine, Zhejiang University.

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Ri, S., Hwang, W., Ri, S. et al. Cloning, characterization, and transcriptional activity of β-actin promoter of African catfish (Clarias gariepinus). Mol Biol Rep 48, 2561–2571 (2021). https://doi.org/10.1007/s11033-021-06306-z

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