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Characterization of mouse myelin basic protein messenger RNAs with a myelin basic protein cDNA clone.

机译:用髓磷脂碱性蛋白cDNA克隆表征小鼠髓磷脂碱性蛋白信使RNA。

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摘要

Using a family of synthetic tetradecamer oligonucleotides as a primer for cDNA synthesis and a second family of tetradecamers as a hybridization probe, we have prepared and isolated a cDNA clone of mouse myelin basic protein (MBP). The clone, pNZ111, corresponds to the region of the mRNA that codes for an amino acid sequence present in all four major forms of MBP. The relative abundance of MBP mRNA, estimated by dot blot hybridization, increased with the age of the mouse to a maximum at 18 days, then decreased to about one-fourth of that amount at later ages. Mouse MBP mRNAs, selected by their ability to hybridize to the clone, translate into the four forms of myelin basic protein. In RNA blot analyses, pNZ111 hybridized to multiple species of mouse mRNA. The predominant hybridization is to a broad band of RNAs ranging in length from 2,350 to 2,100 bases. These mRNA species are extremely long, considering that the largest MBP could be encoded by approximately 600 bases. In addition to these, there are also minor bands that hybridize with pNZ111, including a band of 4,100 bases and smaller ones of 1,900, 1,500, and 1,200 bases.
机译:使用一个合成的四聚体寡核苷酸家族作为cDNA合成的引物,并使用第二个四聚体家族作为杂交探针,我们制备并分离了小鼠髓鞘碱性蛋白(MBP)的cDNA克隆。克隆pNZ111对应于mRNA的区域,该区域编码所有四种主要形式的MBP中存在的氨基酸序列。通过点印迹杂交估计的MBP mRNA的相对丰度随着小鼠的年龄增加而增加,在18天时达到最大值,然后在以后的年龄时降低到其数量的约四分之一。通过小鼠MBP mRNA与克隆杂交的能力对其进行选择,从而将其翻译为四种形式的髓鞘碱性蛋白。在RNA印迹分析中,pNZ111与多种小鼠mRNA杂交。主要的杂交是与宽范围为2350至2100个碱基的RNA杂交。考虑到最大的MBP可以由大约600个碱基编码,因此这些mRNA种类非常长。除这些以外,还有与pNZ111杂交的较小条带,包括4,100个碱基的条带和1,900、1,500和1,200个碱基的较小条带。

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