Roach A, Boylan K, Horvath S, Prusiner S B, Hood L E
Cell. 1983 Oct;34(3):799-806. doi: 10.1016/0092-8674(83)90536-6.
A cDNA library was constructed from mRNA isolated from the brains of 18-day-old rats, the age at which myelin biosynthesis is maximal. A synthetic DNA probe synthesized based on reverse translation of the amino acid sequence of rat myelin basic protein (MBP) was used to select two cDNA clones encoding MBP. A 1.5 kb Eco RI fragment from one clone was completely sequenced. When translated, a portion of this sequence was identical at 126 of 127 positions with the reported amino acid sequence for small MBP from the rat. Brains from mice of the homozygous shiverer genotype contained neatly reduced amounts of MBP mRNA relative to wild type. A deletion of MBP sequences in the genome of shiverer mice was also demonstrated. cDNAs for MBP will allow molecular investigation of the role this gene plays in both dysmyelinating and demyelinating diseases, as well as questions of MBP biosynthesis.
从18日龄大鼠(此时髓鞘生物合成量最大)的大脑中分离出mRNA构建了一个cDNA文库。基于大鼠髓鞘碱性蛋白(MBP)氨基酸序列的反向翻译合成了一种合成DNA探针,用于筛选两个编码MBP的cDNA克隆。对其中一个克隆的1.5 kb Eco RI片段进行了全序列测定。该序列翻译后,在127个位置中的126个位置上与报道的大鼠小MBP氨基酸序列完全相同。相对于野生型,纯合颤抖鼠基因型小鼠的大脑中MBP mRNA含量明显减少。还证实了颤抖鼠小鼠基因组中MBP序列的缺失。MBP的cDNA将有助于对该基因在脱髓鞘疾病和髓鞘破坏疾病中的作用以及MBP生物合成问题进行分子研究。