Zajdel-Blair M E, Blair G E, Bennich H
Nucleic Acids Res. 1981 Sep 25;9(18):4547-55. doi: 10.1093/nar/9.18.4547.
The isolation and translational properties of rat immunoglobulin E (IgE) heavy chain mRNA are described. The mRNA has a sedimentation coefficient of approximately 18S, a chain length of about 2000 nucleotides and directs the synthesis in vitro of a polypeptide of 65000 molecular weight in an mRNA-dependent rabbit reticulocyte lysate. Inclusion of dog pancreatic microsomes in the cell-free translation system resulted in a heavy chain product of about 75000 molecular weight, presumably as a consequence of glycosylation in vitro. This species co-migrated in an SDS polyacrylamide gel with mature IgE heavy chain. Substantial purification of heavy chain mRNA was achieved by denaturing sucrose gradient centrifugation and agarose gel electrophoresis.
本文描述了大鼠免疫球蛋白E（IgE）重链mRNA的分离及其翻译特性。该mRNA的沉降系数约为18S，链长约2000个核苷酸，并在依赖mRNA的兔网织红细胞裂解物中指导体外合成分子量为65000的多肽。在无细胞翻译系统中加入犬胰腺微粒体，产生了分子量约为75000的重链产物，推测这是体外糖基化的结果。该产物在SDS聚丙烯酰胺凝胶中与成熟的IgE重链共同迁移。通过变性蔗糖梯度离心和琼脂糖凝胶电泳实现了重链mRNA的大量纯化。
