Hasan Lara, Vögeli Peter, Stoll Peter, Kramer Spela Spilar, Stranzinger Gerald, Neuenschwander Stefan
Institute of Animal Sciences, Tannenstrasse 1, ETH-Zentrum, CH-8092 Zurich, Switzerland.
Mamm Genome. 2004 Apr;15(4):323-33. doi: 10.1007/s00335-003-2324-6.
The absence of L-ascorbic acid (L-AA, or AA) synthesis in scurvy-prone organisms, including humans, other primates, guinea pigs, and flying mammals, was traced to the lack of L-gulonolactone oxidase (GULO) activity. GULO is a microsomal enzyme that catalyzes the terminal step in the biosynthesis of L-AA. Clinical cases of scurvy were described in a family of Danish pigs. This trait is controlled by a single autosomal recessive allele designated od (osteogenic disorder). Here we demonstrate that the absence of GULO activity and the associated vitamin C deficiency in od/od pigs is due to the occurrence of a 4.2-kbp deletion in the GULO gene. This deletion includes 77 bp of exon VIII, 398 bp of intron 7 and 3.7 kbp of intron 8, which leads to a frame shift. The mutant protein is truncated to 356 amino acids, but only the first 236 amino acids are identical to the wild-type GULO protein. In addition, the od allele seems to be less expressed in deficient and heterozygous pigs compared with the normal allele in heterozygous and wild-type animals as determined by ribonuclease protection assay. We also developed a DNA-based test for the diagnosis of the deficient allele. However, we failed to identify the mutated allele in other pig populations.
在易患坏血病的生物中,包括人类、其他灵长类动物、豚鼠和飞行哺乳动物,缺乏L-抗坏血酸(L-AA,或AA)合成的原因可追溯到缺乏L-古洛糖酸内酯氧化酶(GULO)活性。GULO是一种微粒体酶,催化L-AA生物合成的最后一步。在一个丹麦猪家族中描述了坏血病的临床病例。这个性状由一个单一的常染色体隐性等位基因od(成骨障碍)控制。在这里,我们证明od/od猪中GULO活性的缺失和相关的维生素C缺乏是由于GULO基因中出现了一个4.2kbp的缺失。这个缺失包括外显子VIII的77bp、内含子7的398bp和内含子8的3.7kbp,这导致了移码。突变蛋白被截短为356个氨基酸,但只有前236个氨基酸与野生型GULO蛋白相同。此外,通过核糖核酸酶保护试验确定,与杂合子和野生型动物中的正常等位基因相比,od等位基因在缺乏和杂合子猪中的表达似乎较少。我们还开发了一种基于DNA的检测方法来诊断缺陷等位基因。然而,我们未能在其他猪群中鉴定出突变等位基因。