中华大蟾蜍TAGLN2 cDNA的克隆与其组织分布

Cloning and Tissue Distribution of TAGLN2 in Bufo gargarizans

为研究蟾蜍旧ufo）基原的中药材中的多肽类有效成分，开展从日本蟾蜍（B．japonicus for-moslAs）皮肤cD—NA质粒文库中筛选蛋白编码基因的工作．由于后续拓展蟾蜍药用价值的研究中需要大量的实验材料，开始以中华大蟾蜍（B．gargarizarts）皮肤第一链cDNA为模板，克隆其相关基因的工作．通过DNA聚合酶链式反应获得编码转胶蛋白-2（transgelin2，TAGLN2）的转录子（GenBank登录号为KF432856）．该cDNA全长1207bp，其5’、3’端非翻译区域和开放阅读框分别为16bp、597bp和594bp，编码由197个氨基酸残基组成的蛋白质，与日本蟾蜍的TAGLN2（GenBank登录号为AGQ03775．1）相比，仅一个氨基酸发生替换，与人的同源性为82％，与其他动物的同源性介于70％～75％．RT—PCR组织分布检测表明TAGLN2在所检测的中华大蟾蜍各器官中均有表达．在肺、肝和肾中表达量较多．这些研究结果为后续中华大蟾蜍TAGLN2的生物学功能研究以及相关药物研发提供基础数据．

In previous investigation, Japanese toad （Bufo japonicus formosus） skin plasmid cDNA library was screened to explore the bioactive polypeptides included in the skin and its secretions. The related cDNAs from Chinese toad （B. gargarizans） is cloned for the further exploration of Bufo. As a result, TA GLN2 cDNA was cloned from Chinese toad skin first strand cDNAs （GenBank accession number. KF432856）. The transcript, which is 1 207 bp, is made up of 16 bp 5＇ untranslated region （UTR）, 594 bp 3＇ UTR and an open reading frame of 597 bp encoding a polypeptide of 197 amino acid residues. In Chinese toad TAGLN2, only one amino acid residue is different from Japanese toad TAGLN2. The similarity is 82% with human TAGLN2 and 70%-75% with other animals. The expression level of TA GLN2 in different tissues using RT-PCR was detected. The expression in lung, liver and kidney was more than in other organs. The current study provides the basic data for the further investigation on the TAGLN2 biological functions and its therapeutic drug de-velopment.