目的建立可视化-跨越式滚环等温扩增技术检测肉制品中鼠肉掺假的分析方法。方法选取鼠的线粒体cytb基因为靶基因,设计并筛选出一对引物,选择9个不同物种的新鲜肌肉组织样本为研究对象,验证跨越式滚环等温扩增技术(saltatory rolling cir...目的建立可视化-跨越式滚环等温扩增技术检测肉制品中鼠肉掺假的分析方法。方法选取鼠的线粒体cytb基因为靶基因,设计并筛选出一对引物,选择9个不同物种的新鲜肌肉组织样本为研究对象,验证跨越式滚环等温扩增技术(saltatory rolling circle isothermal amplification, SRCA)方法的特异性;测定该技术的灵敏度以及人工污染样品中鼠肉成分的检出限,验证SRCA方法的准确性。结果 SRCA荧光可视化法检测鼠肉DNA的灵敏度为7.3×100fg/μL,与传统PCR方法相比,SRCA荧光可视化法的灵敏度提高了1000倍。在人工添加鼠肉的模拟掺假样品中, SRCA方法可检测到含量为0.01%的鼠肉。结论 SRCA技术可以灵敏、快速、准确地检测出肉制品中的鼠肉掺假成分,适用于基层单位对肉制品掺假的快速检测。展开更多
This study compared proteome profiles and morphological changes of rat jejunum in response to different dietary proteins. Fifty male Sprague-Dawley rats were fed with casein(control), and isolated beef, pork, fish and...This study compared proteome profiles and morphological changes of rat jejunum in response to different dietary proteins. Fifty male Sprague-Dawley rats were fed with casein(control), and isolated beef, pork, fish and chicken proteins for 14 days. Proteome analysis,histological observation and PEPT1 quantification of the jejunum were performed. The results indicated that rats fed with chicken proteins had higher PEPT1 mRNA and protein levels(P < 0.05) but lower villus height and ratio of villus height to crypt depth(V/C ratio, P < 0.05) than those fed with casein and pork protein. Label-free LC-MS/MS indicated that, as compared to casein, intake of chicken protein can regulate oligopeptide transport mainly by upregulating PEPT1 protein expression and reducing dipeptidyl-peptidase activity related to biological oxidation, and can reduce oligopeptide absorption capacity by regulating Hippo signaling pathway. Although intake of beef and fish proteins had no significant effect on PEPT1 expression, they altered several signaling pathways.展开更多
文摘目的建立可视化-跨越式滚环等温扩增技术检测肉制品中鼠肉掺假的分析方法。方法选取鼠的线粒体cytb基因为靶基因,设计并筛选出一对引物,选择9个不同物种的新鲜肌肉组织样本为研究对象,验证跨越式滚环等温扩增技术(saltatory rolling circle isothermal amplification, SRCA)方法的特异性;测定该技术的灵敏度以及人工污染样品中鼠肉成分的检出限,验证SRCA方法的准确性。结果 SRCA荧光可视化法检测鼠肉DNA的灵敏度为7.3×100fg/μL,与传统PCR方法相比,SRCA荧光可视化法的灵敏度提高了1000倍。在人工添加鼠肉的模拟掺假样品中, SRCA方法可检测到含量为0.01%的鼠肉。结论 SRCA技术可以灵敏、快速、准确地检测出肉制品中的鼠肉掺假成分,适用于基层单位对肉制品掺假的快速检测。
基金supported by the National Natural Science Foundation of China (31471600, 31530054)
文摘This study compared proteome profiles and morphological changes of rat jejunum in response to different dietary proteins. Fifty male Sprague-Dawley rats were fed with casein(control), and isolated beef, pork, fish and chicken proteins for 14 days. Proteome analysis,histological observation and PEPT1 quantification of the jejunum were performed. The results indicated that rats fed with chicken proteins had higher PEPT1 mRNA and protein levels(P < 0.05) but lower villus height and ratio of villus height to crypt depth(V/C ratio, P < 0.05) than those fed with casein and pork protein. Label-free LC-MS/MS indicated that, as compared to casein, intake of chicken protein can regulate oligopeptide transport mainly by upregulating PEPT1 protein expression and reducing dipeptidyl-peptidase activity related to biological oxidation, and can reduce oligopeptide absorption capacity by regulating Hippo signaling pathway. Although intake of beef and fish proteins had no significant effect on PEPT1 expression, they altered several signaling pathways.