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cDNA cloning and characterization of 5′ upstream promoter region of porcine mef2c gene

Molecular Biology Reports volume 38, pages 4723–4730 (2011)Cite this article

Abstract

The myocyte enhancer factor 2C (MEF2C) is a member of the MEF2 family of transcription factors, involved in skeletal muscle development. In this study we report the cDNA sequence and isolate the 5′ upstream region of the mef2c gene from porcine genomic DNA using PCR-based GenomeWalker. The open reading frame of porcine mef2c cDNA covers 1,392 bases, encoding 464 amino acids, which show 94% identity with human MEF2C at the level of the primary protein structure. Annear the C terminus of mef2c, a 96-nt sequence appear to represent alternatively spliced transcripts was present in some cDNAs and absent in the other. No typical TATA, GC box or CAAT box binding site was found in porcine mef2c 5′ upstream region, whereas some potential binding sites for MyoD (E-box), MEF2 and MBF1 were present in the proximal upstream region. Transfection of the mef2c 5′ upstream region with EGFP into cos7 cells demonstrated that the region from –162 to +115 bp immediately 5′ of the exon 1 was sufficient to direct strong EGFP protein expression. Co-transfection assays demonstrated that MBF1 bound the mef2c promoter and inhibited mef2c expression. These results may be useful for elucidating the regulation mechanisms of mef2c, which interacts with other factors to regulate target genes.

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Acknowledgments

The project was supported by the Specialized Research Fund for the Doctoral Program of Higher Education of China (Grant No. 20090146120030 & 20070504064), Natural Science Foundation of Hubei Province of China (Grant No. 2008CDB247), Key Program of National transgenic research of China (Grant No. 2009ZX08009-161B & 2009ZX08009-151B), Key Technologies R&D Program of China (Grant No. 2008BADB2B02-14 & 2006BAD01A08-07), National High-Tech R&D Program of China (Grant No. 2006AA10Z140) and Creative R&D Program of Agricultural Science and Technology of Hubei Province of China (Grant No. 2007-620).

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Affiliations

  1. Agriculture Ministry Key Laboratory of Swine Genetics and Breeding, Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan, People’s Republic of China

    Min Liu, De Quan Xu, Jian Peng, Rong Zheng, Fenge Li & Si Wen Jiang

  2. College of Animal Science and Technology, Huazhong Agricultural University, Wuhan, 430070, People’s Republic of China

    Si Wen Jiang

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  1. Min Liu
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  2. De Quan Xu
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  3. Jian Peng
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  4. Rong Zheng
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  5. Fenge Li
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Correspondence to Si Wen Jiang.

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The authors Min Liu and De Quan Xu contributed equally to this work.

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Liu, M., Xu, D.Q., Peng, J. et al. cDNA cloning and characterization of 5′ upstream promoter region of porcine mef2c gene. Mol Biol Rep 38, 4723–4730 (2011). https://doi.org/10.1007/s11033-010-0609-x

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  • DOI: https://doi.org/10.1007/s11033-010-0609-x

Keywords

  • mef2c
  • Porcine
  • cDNA cloning
  • Promoter analysis
  • Transcription regulation
  • MBF1