荷斯坦奶牛妊娠早期和产后期血浆乳汁差异蛋白质组学研究

发布时间：2018-05-10 11:21

本文选题：荷斯坦奶牛 + 妊娠早期　；参考：《甘肃农业大学》2015年硕士论文

【摘要】：本研究采用二维凝胶电泳(2-DE)结合MADIL-TOF-TOF串联质谱法对荷斯坦奶牛妊娠早期血浆进行蛋白质组学分析鉴定,发现了6种差异蛋白在妊娠后表达量上调,分别是凝集素1前体、转甲状腺素蛋白前体、3,5-二碘水杨酸络合的牛血清白蛋白的晶体结构的A链、白蛋白、未命名蛋白产物及结合珠蛋白-25同源物1前体。凝集素1前体的表达量在发情及配种后呈现先升高后下降的趋势;转甲状腺素蛋白前体,白蛋白的表达量在发情及配种后呈先升高后下降再升高的趋势;3,5-二碘水杨酸络合的牛血清白蛋白的晶体结构的A链的表达量不断增加,未命名蛋白产物、结合珠蛋白-25同源物1前体的含量是先升高后保持稳定。妊娠期这6种蛋白的含量均高于妊娠前,这些蛋白在妊娠早期血浆中可能与免疫应答、甲状腺激素转运、防止淀粉样纤维形成和碘的运输有关。采用相同方法对荷斯坦奶牛分娩后不同时期的血浆进行蛋白质组学分析鉴定,发现5种差异蛋白。结合珠蛋白前体、结合珠蛋白在分娩后表达量先升高后下降至与分娩当天相近。免疫球蛋白γ2链C区(克隆32.2)在分娩后表达量先下降再升高之后保持一个平稳的趋势。分娩后3,5-二碘水杨酸络合的牛血清白蛋白的晶体结构的A链和B链表达量低于妊娠后45天的含量且在分娩后表达量相对稳定。这5种蛋白可能分别与产后应激、免疫应答和物质运输有关。采用二维凝胶电泳(2-DE)结合MADIL-TOF-TOF串联质谱法对荷斯坦奶牛妊娠早期乳汁进行蛋白质组学分析鉴定,发现5个差异蛋白点,分别是:3,5-二碘水杨酸络合的牛血清白蛋白的晶体结构的A链、免疫球蛋白γ1重链恒定区、免疫球蛋白G1重链恒定区、白蛋白、蛋白胨成分3。3,5-二碘水杨酸络合的牛血清白蛋白的晶体结构的A链在乳汁中含量的变化趋势为先降低再升高后降低。免疫球蛋白G1重链恒定区和免疫球蛋白γ1重链恒定区表达量显著降低,之后基本保持稳定的趋势。蛋白胨3的表达量虽有波动,但是妊娠早期高于发情当天。这些蛋白可能与物质运输、免疫应答和抗菌有关。采用相同方法对荷斯坦奶牛分娩后不同时期乳汁进行蛋白质组学分析鉴定,发现8种差异蛋白,分别是:α-S2酪蛋白前体、β-酪蛋白、κ-酪蛋白前体、免疫球蛋白重链前体、白蛋白、血清白蛋白前体、cGMP磷酸二酯酶的α亚基、角蛋白I型细胞骨架10。α-S2酪蛋白前体只在分娩当天乳样中检测到,β-酪蛋白在分娩后7天和分娩后35天检测到,且在分娩后35天的表达量比分娩后7天高,κ-酪蛋白前体在分娩当天表达量最高,之后表达量快速下降之后保持平稳。免疫球蛋白重链前体存在于分娩后7、15、28及35天的乳样中,虽有波动,但在分娩后35天的乳样中表达量最低。血清白蛋白前体在分娩后7天到分娩后35天表达量表现出先升高后降低的趋势,白蛋白在分娩后7天到分娩后35天的表达量呈先降低后升高再降低的趋势,分娩后35天的表达量最低。cGMP磷酸二酯酶的α亚基在分娩后7、15及28天的表达量呈先上升后下降的趋势。角蛋白I型细胞骨架10在分娩当天到分娩后28天表达量的变化趋势平稳。这些蛋白可能与初乳的被动免疫、物质运输和产奶有关。
[Abstract]:In this study, two dimensional gel electrophoresis (2-DE) combined with MADIL-TOF-TOF tandem mass spectrometry was used to identify the proteomic analysis of plasma in the early pregnancy of Holstein cows. It was found that the expression of 6 different proteins increased after pregnancy, which were the 1 agglutinin precursor, the thyroxine precursor, and the 3,5- two iodo salicylic acid complexed bovine serum albumin. The A chain of crystal structure, albumin, unnamed protein products and 1 precursors of binding globin -25 homology. The expression of the 1 agglutinin precursor showed a tendency to rise first and then decline after the estrus and mating; the expression of the HP precursor was increased first and then decreased after the oestrus and distribution; 3,5- two iodide The expression of A chain in the crystal structure of acid complex bovine serum albumin is increasing, the content of the unnamed protein product, the content of the 1 precursor of the binding globin -25 homologue is first and then stable. The content of these 6 proteins in pregnancy is higher than that before pregnancy. These proteins may be associated with the immune response and thyroid hormone transport in the plasma in the early pregnancy. To prevent the formation of amyloid fiber from the transport of iodine. The same method was used to identify the plasma of Holstein cows at different periods after childbirth. 5 differential proteins were found. Combined with globin precursor, the expression of combined globin decreased to the same day after childbirth to the day of childbirth. The 2 chain C area of immunoglobulin gamma (clone 32.2) the expression of 3,5- two iodialicylic acid complex after labor after childbirth is a stable trend. The A chain and B chain expression of the crystal structure of bovine serum albumin complex after delivery are lower than 45 days after pregnancy and are relatively stable after delivery. These 5 proteins may be respectively associated with postpartum stress and immune response, respectively. A two-dimensional gel electrophoresis (2-DE) combined with MADIL-TOF-TOF tandem mass spectrometry was used to identify the proteomic analysis of the early pregnancy milk of Holstein cows. 5 differences were found: the A chain of the crystalline structure of bovine serum albumin complex with 3,5- two iodosalicylic acid, the immunoglobulin gamma 1 heavy chain constant region, and the immunoglobulin gamma. Pestis G1 heavy chain constant region, albumin, peptone composition 3.3,5- two iodosalicylic acid complexed with bovine serum albumin, the crystal structure of bovine serum albumin, the change trend of the content of A chain in milk was decreased first and then decreased. The expression of immunoglobulin G1 heavy chain constant region and immunoglobulin gamma heavy chain constant region decreased significantly, then basically maintained Stable trend. Although the expression of peptone 3 is fluctuating, but early pregnancy is higher than that on the day of estrus. These proteins may be related to material transport, immune response and antibacterial. The same method was used to identify the milk of Holstein cows at different periods after childbirth. 8 different proteins were found, respectively: alpha -S2 casein. Body, beta casein, kappa casein precursor, immunoglobulin heavy chain precursor, albumin, serum albumin precursor, cGMP phosphodiesterase alpha subunit, keratin I cytoskeleton 10. alpha -S2 casein precursor only detected in the milk sample on the day of childbirth, beta casein was detected at 7 days after delivery and 35 days after childbirth, and at 35 days after delivery The highest expression of kappa casein precursor on the day of childbirth was higher than that on the 7 day after delivery, and then the expression level remained stable after the rapid decline. The immunoglobulin heavy chain precursor existed in the 7,15,28 and 35 days of milk after delivery, although there was a fluctuation, but the lowest expression in the milk samples at 35 days after childbirth. The serum albumin precursor was 7 days after childbirth to 3 after childbirth. The expression of the 5 day expression showed a tendency to increase first and then decrease. The expression of albumin at 7 days after delivery to 35 days after childbirth decreased first and then decreased, and the expression of the lowest expression of.CGMP phosphodiesterase at 35 days after childbirth increased first and then declined at 7,15 and 28 days after delivery. The cytoskeleton of keratin I type 10 there was a steady trend in the expression of protein on the day of delivery and 28 days after delivery. These proteins may be related to passive immunity, substance transport and milk production.

【学位授予单位】：甘肃农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S823

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