目的制备牛磷酸烯醇式丙酮酸羧激酶(PEPCK)单克隆抗体(m Ab),并对其特异性进行初步鉴定。方法构建含PEPCK基因的重组质粒,将其转化大肠杆菌,并进行诱导表达,对表达产物PEPCK重组蛋白进行纯化,以PEPCK重组蛋白为抗原,免疫BALB/c小鼠,取脾细胞与小鼠骨髓瘤细胞融合获得杂交瘤细胞,用间接ELISA筛选阳性克隆,获得稳定分泌抗牛PEPCK m Ab的细胞株,并用Western blot法,免疫组织化学染色和免疫沉淀鉴定其特异性。结果成功获得稳定分泌抗牛PEPCK m Ab的杂交瘤细胞株。Western blot法、免疫组织化学和免疫沉淀结果表明,该抗体能与牛PEPCK蛋白特异性结合。结论成功获得抗牛PEPCK m Ab。
Dietary meats play a crucial role in human health. The objective of this survey was to determine the fatty acid content and omega-6/omega-3 polyunsaturated fatty acids （n-6/n-3 PUFA） ratio of fresh red meat （beef and pork） from four cities （Shanghai, Nanjing, Yinchuan and Hohhot） in China. The results showed that the n-6/n-3 PUFA ratio from all the samples ranged from 6 to 23. The total n-6 PUFA concentrations ranged from 290.54 mg/100 g in beef from Nanjing to 1601.48 mg/100 g in pork from Hohhot, whereas the total concentrations of n-3 PUFA ranged from 46.34 mg/100 g in beef from Nanjing to 96.03 mg/100 g in pork from Nanjing. The results indicated that the n-6/n-3 ratio in the red meat from all four regions is unbalanced and is much higher than that （〈 5：1） rec- ommended by the WHO/FAO. The total amount of n-3 PUFA was far lower than the required daily dose. There- fore, potential solutions to increase the n-3 PUFA content in meat products or to provide alternative source of n-3 PUFA should be explored.
Ming YuQianqian GaoYan WangWei ZhangLin LiYing WangYifan Dai
