Cloning and its bioinformatic analysis of <i>EDF-</i>1 cDNA from two<i> Bufo</i> species

FANG Qilu; YUE Jiping; YANG Xianyu

doi:10.11665/j.issn.1000-5048.20140417

Journal of China Pharmaceutical University > 2014 > 45(4): 480-485. > DOI: 10.11665/j.issn.1000-5048.20140417

FANG Qilu, YUE Jiping, YANG Xianyu. Cloning and its bioinformatic analysis of EDF-1 cDNA from two Bufo species[J]. Journal of China Pharmaceutical University, 2014, 45(4): 480-485. DOI: 10.11665/j.issn.1000-5048.20140417

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Cloning and its bioinformatic analysis of EDF-1 cDNA from two Bufo species

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In traditional Chinese medicine, toad skin-origin materials including toad skin and its secretions have long been used to treat inflammation, sores and even many kinds of cancers. To clarify the polypeptides included in these toad-origin materials, screening of cDNAs encoding polypeptides was carried out from the plasmid skin cDNA library of Japanese toad(Bufo japonicus formosus)by colony polymerase chain reaction(PCR). As the result, a transcript of 958 bp(GenBank accession number: KF769458)was obtained consisting of 13 bp 5′ untranslated region(UTR), 498 bp 3′ UTR and an open reading frame(ORF)of 447 bp encoding a polypeptide of 148 amino acid residues with high homology with endothelial differentiation-related factor-1(EDF-1)of other animals. Based on the EDF-1 cDNA sequence of Japanese toad, a pair of primers were designed for cloning Chinese toad(B. gargarizans) EDF-1 and a DNA segment was obtained with an ORF of 447 bp(GenBank accession number: KF769459)encoding a polypeptide of 148 amino acids, among which three amino acid residues were different from that of Japanese toad. For two Bufo species, the similarity is 95% with human EDF-1 and 93%-97% with other animals indicating that EDF-1 is evolutionarily highly conserved. Besides, phosphorylation site prediction indicated 9 potential sites suggesting its activity being upstream regulated which is consistent with previous reports. Due to the involement in the process of endothelial cell differentiation, EDF-1 expressed in toad skin might be one of the important ingredients with antitumor effects.
- Bufo japonicus formosus,
- B.gargarizans,
- EDF-1,
- gene cloning,
- bioinformatic analysis

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