Abstract
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, TiO2 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.
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Acknowledgements
This work was supported by the National Transgenic Animal Program (Grant Nos. 2014ZX08007-002, 2016ZX08007002-004) of China. This work was accomplished in the Tianjin International Joint Research and Development Center. We thank The Key Laboratory of Mammalian Reproductive Biology and Biotechnology of the Ministry of Education, Inner Mongolia University, for proving us with experimental materials. We also want to thank the Proteomic and MS Facility of Cornell University and the team of Professor Sheng Zhang for generating the mass spectrometry data and NIH SIG 1S10 OD017992-01 grant support for the Orbitrap Fusion mass spectrometer. We are grateful to the help of Agricultural animal breeding and product testing Biotechnology Research Center of Tianjin Agricultural University.
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HG, XFL, GPL and XBX conceived and designed the experiments. XBX and SPY performed most of the experiments, and XBX performed the data analyses and wrote the manuscript. CFJ and SPY contributed significantly to performed animal samples preparation. XL, XBD and XFL contributed significantly to revise the manuscript. All authors read and approved the final manuscript.
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Xin, X., Liu, X., Li, X. et al. Comparative muscle proteomics/phosphoproteomics analysis provides new insight for the biosafety evaluation of fat-1 transgenic cattle. Transgenic Res 26, 625–638 (2017). https://doi.org/10.1007/s11248-017-0032-3
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DOI: https://doi.org/10.1007/s11248-017-0032-3