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Molecular cloning, prokaryotic expression and promoter analysis of squalene synthase gene from Schizochytrium Limacinum

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Abstract

Squalene synthase (SQS) is an important enzyme in the steroid biosynthetic pathways which condenses two molecules of farnesyl pyrophosphate into a squalene. In this study, the gene encoding SQS was isolated from Schizochytrium limacinum and characterized. The full-length cDNA of S. limacinum SQS gene (SlSQS) is 1605 bp in length, it contains a 1293 bp ORF encoding a polypeptide of 430 amino acids. Multiple amino acid sequence alignment showed that the SlSQS protein sequence shared 5 conserved signature domains and a hydrophobic carboxy-terminal part with other known SQS protein sequences. C-terminal-truncated SlSQS was constructed into expression vector pGEX and successfully expressed in Escherichia coli cells. The expressed fusion protein was confirmed to have SQS activity. In addition, a 724 bp promoter region of SlSQS was also cloned and several cis-acting elements were predicted. These results might be helpful to understand the structure and expression regulation of SQS in S. limacinum.

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Authors and Affiliations

  1. College of Agriculture, Ludong University, Yantai, 264025, China

    L. Zhu, L. Chang, P. Feng & L. Han

  2. College of Marine Life Sciences, Ocean University of China, Qingdao, 266003, China

    X. Zhang

  3. College of Life Sciences, Ludong University, Yantai, 264025, China

    A. Wang

Authors
  1. L. Zhu
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  2. X. Zhang
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Correspondence to L. Zhu.

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Zhu, L., Zhang, X., Chang, L. et al. Molecular cloning, prokaryotic expression and promoter analysis of squalene synthase gene from Schizochytrium Limacinum . Appl Biochem Microbiol 50, 411–419 (2014). https://doi.org/10.1134/S0003683814040140

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  • DOI: https://doi.org/10.1134/S0003683814040140

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