College of Animal Science and Veterinary Medicine, Tianjin Agriculture University, 22 Jinjing Road, Tianjin, 300384, China.
The Key Laboratory of Mammalian Reproductive Biology and Biotechnology of the Ministry of Education, Inner Mongolia University, 24 Zhaojun Road, Hohhot, 010070, China.
Transgenic Res. 2017 Oct;26(5):625-638. doi: 10.1007/s11248-017-0032-3. Epub 2017 Jul 14.
The biosafety of fat-1 transgenic cattle has been a focus of our studies since the first fat-1 transgenic cow was born. In this study, we used tandem mass tag labeling, TiO enrichment, and nanoscale liquid chromatography coupled with tandem mass spectrometry (nanol LC-MS/MS) to compare proteomic and phosphoproteomic profiling analyses of muscle between fat-1 transgenic cows and wild-type cows. A total of 1555 proteins and 900 phosphorylation sites in 159 phosphoproteins were identified in the profiling assessments, but only four differentially expressed proteins and nine differentially expressed phosphopeptides were detected in fat-1 transgenic cows relative to wild-type cows. Bioinformatics analyses showed that all of the identified proteins and phosphoproteins were mainly related to the metabolic processes of three major nutrients: carbohydrates, lipids, and proteins. All of these differentially expressed proteins might take part in DNA recombination, repair, and regulation of the immune system. In conclusion, most of the identified proteins and phosphoproteins exhibited few changes. Our results provide new insights into the biosafety of fat-1 transgenic cattle.
自第一头 fat-1 转基因奶牛诞生以来,其生物安全性一直是我们研究的重点。在这项研究中,我们使用串联质量标签标记、TiO2 富集和纳升液相色谱与串联质谱(nano LC-MS/MS)技术,比较了 fat-1 转基因奶牛和野生型奶牛肌肉的蛋白质组学和磷酸化蛋白质组学分析。在分析中鉴定出了 1555 种蛋白质和 159 种磷酸化蛋白质中的 900 个磷酸化位点,但在 fat-1 转基因奶牛与野生型奶牛的比较中,只检测到 4 种差异表达蛋白和 9 种差异表达的磷酸肽。生物信息学分析表明,所有鉴定出的蛋白质和磷酸化蛋白质主要与三大营养素(碳水化合物、脂肪和蛋白质)的代谢过程有关。所有这些差异表达的蛋白质可能参与 DNA 重组、修复和免疫系统的调节。总之,大多数鉴定出的蛋白质和磷酸化蛋白质变化不大。我们的研究结果为 fat-1 转基因奶牛的生物安全性提供了新的见解。