Farrell L B, Beachy R N
Washington University, Department of Biology, St. Louis, MO 63130.
Plant Mol Biol. 1990 Dec;15(6):821-5. doi: 10.1007/BF00039422.
It has been documented that when furnished with an endomembrane signal sequence for the endoplasmic reticulum, beta-glucuronidase (GUS) is N-glycosylated, resulting in the nearly complete loss of enzymatic activity. To enable use of beta-glucuronidase as a reporter protein in secretory and vacuolar targeting studies, one of the two putative N-linked glycosylation sites within the GUS gene was altered by site-directed mutagenesis. The second N-linked glycosylation site was not altered because sequence analysis of nucleotide sequences around the second putative glycosylation site revealed that the published sequence was incorrect, and that no such site existed.
据记载，当为β-葡萄糖醛酸酶（GUS）提供内质网内膜信号序列时，它会进行N-糖基化，导致酶活性几乎完全丧失。为了能够在分泌和液泡靶向研究中使用β-葡萄糖醛酸酶作为报告蛋白，通过定点诱变改变了GUS基因内两个假定的N-连接糖基化位点之一。第二个N-连接糖基化位点未改变，因为对第二个假定糖基化位点周围核苷酸序列的分析表明，已发表的序列是错误的，不存在这样的位点。
