Characterization of mouse myelin basic protein messenger RNAs with a myelin basic protein cDNA clone.
Characterization of mouse myelin basic protein messenger RNAs with a myelin basic protein cDNA clone.
复制标题
使用髓磷脂碱性蛋白 cDNA 克隆表征小鼠髓磷脂碱性蛋白信使 RNA。
DOI：
10.1073/pnas.81.1.18
复制
发表时间：
1984
影响因子：
11.1
通讯作者：
Lazzarini,RA
中科院分区：
文献类型：
--
作者：
Zeller,NK;Hunkeler,MJ;Campagnoni,AT;Sprague,J;Lazzarini,RA
使用合成的十四聚体寡核苷酸家族作为 cDNA 合成的引物，并使用第二个十四聚体家族作为杂交探针，我们制备并分离了小鼠髓磷脂碱性蛋白（MBP）的 cDNA 克隆。该克隆 pNZ111 对应于编码 MBP 所有四种主要形式中存在的氨基酸序列的 mRNA 区域。通过斑点印迹杂交估计，MBP mRNA 的相对丰度随着小鼠年龄的增长而增加，在 18 天时达到最大值，然后在以后的年龄中减少到该数量的约四分之一。小鼠 MBP mRNA 根据其与克隆杂交的能力进行选择，并翻译成四种形式的髓磷脂碱性蛋白。在 RNA 印迹分析中，pNZ111 与多种小鼠 mRNA 杂交。主要杂交是与长度范围为 2,350 至 2,100 个碱基的宽带 RNA 杂交。这些 mRNA 种类非常长，考虑到最大的 MBP 可以由大约 600 个碱基编码。除此之外，还有一些与 pNZ111 杂交的小条带，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。
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.