Cloning the sterol carrier protein 2 genes of Japanese toad (Bufo japonicus formosus) and Chinese toad (Bufo gargarizans) and its tissue expression analysis

Dongwuxue Yanjiu. 2014 Sep;35(5):398-403. doi: 10.13918/j.issn.2095-8137.2014.5.398.

Abstract

In this study, to clarify the bioactive polypeptides included in the skins and secretions of Bufo, we screened the Japanese toad (Bufo japonicus formosus) skin cDNA liary by colony polymerase chain reaction (PCR), and obtained a transcript of 1 075 bp consisting of 1 37 bp 5' untranslated region (UTR), 515 bp 3' UTR and a 423 bp open reading frame (ORF) encoding a polypeptide of 140 amino acid residues (GenBank accession number: KF359945). Homolog analysis showed a 70%-96% homology with sterol carrier protein-2 (SCP-2) present in other animals, which is implicated in lipid metabolism of other organisms. The gene SCP-2 of Chinese toad (B. gargarizans) was cloned from a first strand cDNA of Bufo skin (GenBank accession number: KF381341) via PCR, whose encoding polypeptide has only one amino acid difference from that of Japanese toad. Tissue distribution analysis showed that SCP-2 expressed in all organs tested, though in the liver and spleen it manifested lower expression than in other organs. These findings might indicate SCP-2 being one of the active ingredients in toad skin. These findings may in turn have implications for further drug development from traditional Chinese medicine sources.

Keywords: Bufo gargarizans; Bufo japonicus formosus; SCP-2 cDNA cloning; Tissue expression.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Bufonidae / genetics
  • Bufonidae / metabolism*
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism*
  • Catalytic Domain
  • Cloning, Molecular*
  • Gene Expression Regulation / physiology*
  • Molecular Sequence Data
  • Phylogeny
  • Species Specificity

Substances

  • Carrier Proteins
  • sterol carrier proteins

Grants and funding

This study was supported by the National Natural Science Foundation of China (31071181, 31372149) and the Students' Innovative Training Program of ZAFU (20120207, 20120213)