中文摘要：

瞄准：在胃肠的系统克隆并且描绘猪的 aquaporins (AQP ) 。方法：基于 PCR 的克隆策略和赛跑被用来克隆从相对地抄录的猪肝 cDNA 编码顺序的全身的 AQP。散布的站流动光和一个基于 YFP 的荧光方法被用来测量红血球的渗透的透水性并且稳定地 transfected CHO 房间。RT-PCR，北污点，和免疫组织化学被用来决定克隆的 AQP 的胃肠的表示和本地化。在 transfected 房间和红细胞的蛋白质表示被西方的污点分析。结果：编码 271 氨基酸的 813 bp cDNA 猪的 aquaporin (指明的 pAQP1 ) 从肝 mRNA 被克隆(pAQP1 与人的 AQP1 有 93% 身份并且包含在 AQP 家庭，保存的二个 NPA 主题为连接 N 的 glycosylation 的一个一致序列，和在半胱氨酸 191 点的一个水银敏感的地点) 。RT-PCR 分析在猪的消化的腺和内脏揭示了 pAQP1 mRNA 的广泛的表示。北污点在选择消化机关给一个单身者看了 3.0 kb 抄本。pAQP1 蛋白质在小肠，唾液的腺的 microvessles，以及由 immunoperoxydase 的肝内胆汁管的上皮的中央 lacteals 是局部性的。是的高渗透的透水性禁止由 HgCl2 能稳定地在猪的红血球和 CHO 房间被检测有 pAQP1 cDNA 的 transfected。猪的红血球和 pAQP-transfected CHO 房间的 Immunoblot 分析揭示了 unglycosylated 28 ku 乐队和更大的 glycosylated 蛋白质。结论：pAQP1 是到目前为止能分子地被识别的第一猪的 aquaporin。在猪的消化机关的上皮和内皮细胞层的 pAQP1 的宽广分发可以在液体分泌物 / 吸收以及在胃肠的系统的消化功能和病理生理学建议调停隧道的水运输的一个重要角色。

英文摘要：

AIM： To clone and characterize the porcine aquaporins （AQPs） in the gastrointestinal system. METHODS： A PCR-based cloning strategy and RACE were used to clone full-length AQP coding sequence from reversely transcribed pig liver cDNA. Stopped-flow light scattering and a YFP-based fluorescence method were used to measure the osmotic water permeability of erythrocytes and the stably transfected CHO cells. RT-PCR, Northern blot, and immunohistochemistry were used to determine the gastrointestinal expression and localization of cloned AQPs. Protein expression in transfected cells and red blood cells was analyzed by Western blot. RESULTS： An 813 bp cDNA encoding a 271 amino acid porcine aquaporin （designated pAQP1） was cloned from liver mRNA （pAQP1 has a 93% identity with human AQP1 and contains two NPA motifs conserved in AQP family, one consensus sequence for N-linked glycosylation, and one mercury-sensitive site at cysteine 191）. RT-PCR analysis revealed extensive expression of pAQP1 mRNA in porcine digestive glands and gut. Northern blot showed a single 3.0 kb transcript in selected digestive organs, pAQP1 protein was localized at central lacteals of the small intestine, microvessles of salivary glands, as well as epithelium of intrahepatic bile ducts by immunoperoxydase. High osmotic water permeability that is inhibitable by HgCI2 was detected in porcine erythrocytes and CHO cells stably transfected with pAQP1 cDNA. Immunoblot analysis of porcine erythrocytes and pAQP-transfected CHO cells revealed an unglycosylated 28 ku band and larger glycosylated proteins. CONCLUSION： pAQP1 is the first porcine aquaporin that can be molecularly identified so far. The broad distribution of pAQP1 in epithelium and endothelium of porcine digestive organs may suggest an important role of channel-mediated water transport in fluid secretion/absorption as well as in digestive function and pathophysiology of the gastrointestinal system.