Gauss Katherine A, Bunger Peggy L, Siemsen Daniel W, Young Catherine J, Nelson-Overton Laura, Prigge Justin R, Swain Steve D, Quinn Mark T
Department of Veterinary Molecular Biology and Center for Bison and Wildlife Health, Montana State University, Bozeman, MT 59717, USA.
Comp Biochem Physiol B Biochem Mol Biol. 2002 Sep;133(1):1-12. doi: 10.1016/s1096-4959(02)00090-8.
During the host defense process, neutrophils migrate into infected tissues where they become activated, resulting in the assembly of a superoxide anion-generating complex known as the NADPH oxidase. Despite the importance of this system in animal host defense, almost nothing is known about the NADPH oxidase in neutrophils from wild ruminant species. In the present studies, we provide a molecular analysis of the bison leukocyte NADPH oxidase. Using reverse transcriptase-polymerase chain reaction and rapid amplification of cDNA ends, we cloned and sequenced the full-length cDNAs for five bison NADPH oxidase components: p22(phox), p40(phox), p47(phox) and p67(phox), and gp91(phox). When compared to other species, the deduced amino acid sequences of the bison homologs were most similar to those of bovine. Interestingly, a bison p40(phox) alternative splice product was isolated, which was similar to that observed for human p40(phox) in that the cDNAs contained sequence from intron 8. Consistent with the high degree of similarity between bison and bovine amino acid sequences, immunoblot analysis showed that the bison homologs migrated similarly to their bovine counterparts. Overall, these studies show that the bison and bovine NADPH oxidase genes are highly conserved between these two species, despite their divergence from a common ancestor over 1 million years ago.
在宿主防御过程中，中性粒细胞迁移到受感染组织并在那里被激活，导致一种称为NADPH氧化酶的超氧阴离子生成复合物的组装。尽管该系统在动物宿主防御中很重要，但对于野生反刍动物物种中性粒细胞中的NADPH氧化酶几乎一无所知。在本研究中，我们对野牛白细胞NADPH氧化酶进行了分子分析。利用逆转录聚合酶链反应和cDNA末端快速扩增技术，我们克隆并测序了野牛NADPH氧化酶五个组分的全长cDNA：p22(phox)、p40(phox)、p47(phox)、p67(phox)和gp91(phox)。与其他物种相比，野牛同源物的推导氨基酸序列与牛的最为相似。有趣的是，分离出了一种野牛p40(phox)可变剪接产物，它与人类p40(phox)中观察到的类似，因为cDNA包含来自内含子8的序列。与野牛和牛氨基酸序列的高度相似性一致，免疫印迹分析表明，野牛同源物的迁移与它们的牛对应物相似。总体而言，这些研究表明，尽管野牛和牛在100多万年前从共同祖先分化而来，但它们的NADPH氧化酶基因在这两个物种之间高度保守。
