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Comparative Proteomics of Milk Fat Globule Membrane Proteins from Transgenic Cloned Cattle

机译:转基因克隆牛乳脂球膜蛋白的比较蛋白质组学

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摘要

The use of transgenic livestock is providing new methods for obtaining pharmaceutically useful proteins. However, the protein expression profiles of the transgenic animals, including expression of milk fat globule membrane (MFGM) proteins, have not been well characterized. In this study, we compared the MFGM protein expression profile of the colostrum and mature milk from three lines of transgenic cloned (TC) cattle, i.e., expressing recombinant human α-lactalbumin (TC-LA), lactoferrin (TC-LF) or lysozyme (TC-LZ) in the mammary gland, with those from cloned non-transgenic (C) and conventionally bred normal animals (N). We identified 1, 225 proteins in milk MFGM, 166 of which were specifically expressed only in the TC-LA group, 265 only in the TC-LF group, and 184 only in the TC-LZ group. There were 43 proteins expressed only in the transgenic cloned animals, but the concentrations of these proteins were below the detection limit of silver staining. Functional analysis also showed that the 43 proteins had no obvious influence on the bovine mammary gland. Quantitative comparison revealed that MFGM proteins were up- or down-regulated more than twofold in the TC and C groups compared to N group: 126 in colostrum and 77 in mature milk of the TC-LA group; 157 in colostrum and 222 in mature milk of the TC-LF group; 49 in colostrum and 98 in mature milk of the TC-LZ group; 98 in colostrum and 132 in mature milk in the C group. These up- and down-regulated proteins in the transgenic animals were not associated with a particular biological function or pathway, which appears that expression of certain exogenous proteins has no general deleterious effects on the cattle mammary gland.
机译:转基因家畜的使用为获得可药用蛋白质提供了新方法。但是,转基因动物的蛋白质表达谱,包括乳脂球膜(MFGM)蛋白的表达,尚未得到很好的表征。在这项研究中,我们比较了转基因克隆(TC)牛的三系,即表达重组人α-乳清蛋白(TC-LA),乳铁蛋白(TC-LF)或溶菌酶的初乳和成熟牛奶的MFGM蛋白表达谱。 (TC-LZ),以及来自克隆的非转基因(C)和常规繁殖的正常动物(N)的乳腺。我们在牛奶MFGM中鉴定了1,225种蛋白质,其中166种仅在TC-LA组中特异性表达,265种仅在TC-LF组中表达,仅184种在TC-LZ组中特异性表达。仅在转基因克隆动物中表达了43种蛋白质,但这些蛋白质的浓度低于银染的检测极限。功能分析还显示43种蛋白质对牛乳腺无明显影响。定量比较显示,TC和C组的MFGM蛋白上调或下调是N组的两倍以上:TC-LA组的初乳中为126,成熟乳中为77; TC-LF组初乳中157人,成熟乳中222人; TC-LZ组的初乳中有49%,成熟乳中有98%; C组初乳中98份,成熟乳中132份。转基因动物中这些上调和下调的蛋白质与特定的生物学功能或途径无关,这似乎表明某些外源蛋白质的表达对牛乳腺没有一般的有害作用。

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