Identification of whale species by thin-layer isoelectric focusing of sarcoplasmic proteins.

Y. Ukishima, M. Kino, H. Kubota, S. Wada, S. Okada
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引用次数: 5

Abstract

Thin-layer isoelectric focusing was applied to the identification of whale (Cetacea) species by using water-soluble sarcoplasmic proteins of skeletal muscles. Twenty-eight samples consisting of 4 species (10 samples) of baleen whales (Mysticeti) and 8 species (18 samples) of toothed whales (Odontoceti) were analyzed. Each sample (approximately 1 g) was electrophoresed with Ampholine PAGplate, pH 3.5-9.5. The electrophoretic profiles were species-specific on the 4 toothed whale species that did not have a marked intra-species difference, and all 4 baleen whale species. However, the profiles were not specific on the 4 other dolphin species, even though they were discriminable from the other 4 toothed whale species. Numerical values of pIs and relative peak heights were obtained by densitometric analysis of the isoelectro-focused protein bands. The bands were also species-specific for the 8 toothed whale species mentioned. The values may be applicable to species identification without the need for a standard sample, which may not be readily obtainable. Experiments on test samples of minke and sel whales showed that bloodletting with ice water made the densities of isoelectro-focused bands thinner, although species identification was still possible by using the inside part of muscles. Heat treatment at below 60 degrees C for 10 min caused little denaturation; at higher temperatures the protein bands were diminished in a temperature-dependent fashion. Therefore, the present isoelectric focusing analysis should be applicable to small samples of whale meat, excluding several species of dolphins.
用肌浆蛋白薄层等电聚焦法鉴定鲸鱼种类。
采用骨骼肌水溶性肌浆蛋白对鲸类进行了薄层等电聚焦鉴定。分析了28个样本,其中须鲸(神秘鲸目)4种(10个样本)和齿鲸(齿鲸目)8种(18个样本)。每个样品(约1g)用ph3.5 -9.5的Ampholine page板电泳。电泳图谱在种内差异不显著的4种齿鲸和4种须鲸中均具有种特异性。然而,这些资料并不是针对其他4种海豚的,尽管它们与其他4种齿鲸是可以区分的。通过等电聚焦蛋白带的密度分析,得到了pi值和相对峰高的数值。这些条带对于提到的8种齿鲸物种也是特定的。这些值可能适用于物种鉴定,而不需要标准样品,而标准样品可能不易获得。对小须鲸和小须鲸的测试样本进行的实验表明,用冰水放血会使等电聚焦带的密度变薄,尽管仍然可以通过使用肌肉的内部部分来识别物种。在60℃以下热处理10min,变性很小;在较高的温度下,蛋白质带以温度依赖的方式减少。因此,目前的等电聚焦分析应该适用于鲸鱼肉的小样本,不包括几种海豚。
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