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首页> 外文期刊>Zoological Science >Characterization of Alpha-fetoprotein in Fetal Striped Dolphin (Stenella coeruleoalba): Purification of Protein Product and Molecular Cloning of the Corresponding Transcript
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Characterization of Alpha-fetoprotein in Fetal Striped Dolphin (Stenella coeruleoalba): Purification of Protein Product and Molecular Cloning of the Corresponding Transcript

机译：胎儿条纹海豚（Stenella coeruleoalba）中甲胎蛋白的表征：蛋白产物的纯化和相应转录本的分子克隆

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摘要

Alpha-fetoprotein (AFP) is a fetal glycoprotein that is known as a biomarker for monitoring pregnancy in many mammalian species. However, characterization of AFP has not yet been undertaken in any cetacean species. Here, we purified AFP from the serum of fetal striped dolphin by chemical precipitation followed by a combination of immunoadsorbent column chromatography and gel filtration. The molecular masses of native and denatured dolphin AFP were estimated to be similar to 78,000 Da by gel filtration and similar to 68,000 Da by SDS-PAGE, respectively, representing typical masses reported for mammalian AFPs. In fetal serum, only the AFP band (similar to 68,000 Da) appeared to be immunoreactive to an antiserum against purified dolphin AFP, indicating sufficient specificity for the development of an AFP immunoassay. Full-length cDNA encoding for the dolphin AFP was cloned from fetal liver and revealed an open reading frame comprising 610 amino acid residues, which included a putative signal peptide of 18 amino acid residues. This was followed by a sequence identical to the N-terminus of purified AFP. The deduced amino acid sequence of dolphin AFP showed more than 80% identity to those of other mammalian AFPs. To our knowledge, the present report represents the first identification and characterization of AFP from any cetacean species.
机译：甲胎蛋白（AFP）是一种胎儿糖蛋白，被称为监测许多哺乳动物物种怀孕的生物标志物。但是，尚未在任何鲸类物种中进行AFP的表征。在这里，我们通过化学沉淀，然后结合免疫吸附柱层析和凝胶过滤，从胎儿条纹海豚的血清中纯化了AFP。天然和变性海豚AFP的分子量通过凝胶过滤估计分别接近78,000 Da，通过SDS-PAGE估计接近68,000 Da，代表了哺乳动物AFP的典型质量。在胎儿血清中，只有AFP条带（约68,000 Da）似乎对针对纯化的海豚AFP的抗血清具有免疫反应性，表明对AFP免疫测定法的开发具有足够的特异性。从胎儿肝脏中克隆了编码海豚AFP的全长cDNA，揭示了一个开放阅读框，其中包含610个氨基酸残基，其中包括一个推测的18个氨基酸残基的信号肽。其后是与纯化的AFP的N端相同的序列。海豚AFP的推导氨基酸序列与其他哺乳动物AFP的同源性超过80％。据我们所知，本报告是对任何鲸类物种中AFP的首次鉴定和表征。

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