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不同保存方法对厚壳贻贝样品蛋白提取效果的比较 |
杨金龙1,2,3, 李无霜1, 梁 箫1, 沈和定1,2,3, 李家乐1,2,3
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1.上海海洋大学 农业部淡水水产种质资源重点实验室;2.上海市水产养殖工程技术研究中心;3.上海高校知识服务平台上海海洋大学水产动物遗传育种中心
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摘要: |
为了解厚壳贻贝(Mytilus coruscus)稚贝样品在冷冻保存过程中的蛋白变化情况,作者采用直接冷冻法、添加海水冷冻法、蛋白裂解液冷冻法等分别对保存1周至6个月的厚壳贻贝样品蛋白提取的不同效果进行了比较。SDS-PAGE胶结果显示,3种冷冻保存方法均出现4个有规律条带(a、b、c、d),其相对分子量依次约为55、46、38 和26kDa。SDS-PAGE 蛋白条带灰度值的检测结果表明,1个月内蛋白裂解液法保存的样品最佳;直接冷冻法和添加海水冷冻法的研究发现保存过程中的蛋白均出现不同程度的降解现象,因而可能不适宜用于该种稚贝样品的保存。 |
关键词: 厚壳贻贝(Mytilus coruscus) 稚贝 SDS-PAGE 电泳 蛋白条带 |
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基金项目:国家自然科学基金资助项目(31101885); 上海市科委重点支撑项目(12230502100) |
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Effects of cryopreservation methods on protein extraction of the juvenile Mytilus coruscus |
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Abstract: |
In the laboratory, three cryopreservation methods including direct cryopreservation, cryopreservation with seawater, cryopreservation with protein lysis buffer were used to compare their effects on protein extraction of juvenile Mytilus coruscus. Proteins of each sample were extracted after being conserved for one week, two weeks, three weeks, four weeks, two months, three months and six months. The results of SDS-PAGE indicated that four protein bands appeared during all the periods using three cryopreservation methods, and their relative molecular weights were 55, 46, 38 and 26 kDa, respectively. The protein band grayscale values of SDS-PAGE in three cryopreservation methods showed that cryopreservation with protein lysis buffer within one month exhibited the best results on protein extraction, indicating that cryopreservation with protein lysis buffer may be the most appropriate during the present investigation on the protein extraction. In contrast, protein degradation was observed during the preservation using direct cryopreservation and cryopreservation with seawater, indicating that these two methods may be not suitable for protein extraction of the juvenile Mytilus coruscus. |
Key words: Mytilus coruscus juvenile SDS-PAGE protein band |