【目的】探讨藏猪仔猪肠道N-乙酰谷氨酸合成酶(NAGS)在哺乳期的表达变化。【方法】克隆藏仔猪的NAGS基因,并制备其多克隆抗体,分别采用实时定量PCR和Western blotting方法检测1、7、21、28和35日龄的哺乳藏仔猪空肠和回肠中NAGS mRNA的表达水平和蛋白丰度。【结果】7~35日龄藏仔猪空肠与回肠的NAGS mRNA表达水平(P〈0.05)和蛋白丰度(P〈0.01)均随日龄增长呈线性下降趋势。【结论】哺乳期仔猪的空肠和回肠NAGS mRNA和蛋白表达水平随日龄增长而下降,可能是内源性精氨酸合成减少的一种重要机制。
Objective This study was conducted to investigate the mechanism for the reduction of endogenous arginine synthesis by profiling the developmental expression pattern of intestinal N-acetylglutamate synthase(NAGS)in pre-weaned Tibetan piglets.Method The gene of NAGS was cloned from Tibetan piglets,and the rabbit antiserum against the purified NAGS was prepared.The gene expression level and protein content of NAGS were analyzed using real time PCR and western blotting methods in jejunum and ileum of Tibetan piglets at 1, 7, 21, 28 and 35 days of age, respectively. [Result] With the age increasing from day 1 to 35, both the mRNA expression level (P〈0.05) and protein content (P〈0.01) of NAGS were gradually decreased. [ Conclusion] The expression levels of NAGS mRNA and protein were decreased during sucking period as the Tibetan piglet grew up. The decreased expression level of NAGS may be one of the important mechanisms for the reduction of endogenous arginine synthesis in pre-weaned suckling piglets.