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日本蟾蜍(Bufo japonicus formosus)和中华蟾蜍(Bufo gargarizans)固醇载体蛋白2基因的克隆及其组织表达分析。

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

作者信息

Ji Yu-Cheng, Zhuge Hui, Zhang Shan-Shan, Zhang Shu-Fang, Yang Xian-Yu

机构信息

School of Forestry and Biotechnology, Zhejiang Agricultural and Forestry University, Lin'an 311300, China.

The Nurturing Station for the State Key Laboratory of Subtropical Silviculture, Zhejiang Agricultural and Forestry University, Lin'an 311300, China.

出版信息

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.

摘要

在本研究中,为了阐明蟾蜍皮肤和分泌物中所含的生物活性多肽,我们通过菌落聚合酶链反应(PCR)筛选了日本蟾蜍(Bufo japonicus formosus)皮肤cDNA文库,并获得了一个1075 bp的转录本,它由137 bp的5'非翻译区(UTR)、515 bp的3'UTR和一个423 bp的开放阅读框(ORF)组成,该开放阅读框编码一个含140个氨基酸残基的多肽(GenBank登录号:KF359945)。同源性分析显示,其与其他动物体内存在的固醇载体蛋白-2(SCP-2)具有70%-96%的同源性,SCP-2与其他生物的脂质代谢有关。中华蟾蜍(B. gargarizans)的SCP-2基因通过PCR从蟾蜍皮肤的第一链cDNA中克隆得到(GenBank登录号:KF381341),其编码的多肽与日本蟾蜍的多肽仅有一个氨基酸差异。组织分布分析表明,SCP-2在所有检测的器官中均有表达,不过在肝脏和脾脏中的表达低于其他器官。这些发现可能表明SCP-2是蟾蜍皮肤中的活性成分之一。这些发现可能进而对从中药来源进一步开发药物具有启示意义。

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