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大鼠免疫球蛋白ε重链的克隆cDNA探针:构建、鉴定及DNA序列

A cloned cDNA probe for rat immunoglobulin epsilon heavy chain: construction, identification, and DNA sequence.

作者信息

Kindsvogel W R, Reddy E P, Moore J M, Faust C H

出版信息

DNA. 1982;1(4):335-43. doi: 10.1089/dna.1982.1.335.

Abstract

IgE has clinical importance because it is responsible for immediate hypersensitivity. Studies of IgE expression in rats appear to contradict current models for immunoglobulin gene expression. To study rat IgE expression at the RNA and DNA levels, we have constructed a cDNA for part of the rat epsilon (epsilon) heavy chain that is expressed by a rat myeloma, IR162. The rat epsilon-chain clone was initially identified by an efficient selection scheme. DNA sequencing of the 580-bp cDNA revealed that it encoded 176 amino acids that were 50% homologous to the human epsilon H chain. The sequence begins near the end of the CH2 domain and ends 31 amino acids into the CH4 domain. Cysteines important for the structure of the human IgE were conserved in the rat epsilon H-chain. The identity of the cloned epsilon cDNA was confirmed by comparison with a portion of the constant region gene for mouse epsilon H chain. The mouse and rat nucleotide sequences were 79% homologous.

摘要

免疫球蛋白E(IgE)具有临床重要性,因为它介导速发型超敏反应。对大鼠体内IgE表达的研究似乎与当前免疫球蛋白基因表达模型相矛盾。为了在RNA和DNA水平上研究大鼠IgE的表达,我们构建了大鼠ε(epsilon)重链部分的cDNA,该重链由大鼠骨髓瘤IR162表达。大鼠ε链克隆最初是通过一种高效筛选方案鉴定出来的。对580个碱基对的cDNA进行DNA测序后发现,它编码176个氨基酸,与人类ε重链有50%的同源性。该序列起始于CH2结构域末端附近,终止于CH4结构域内31个氨基酸处。对人类IgE结构重要的半胱氨酸在大鼠ε重链中是保守的。通过与小鼠ε重链恒定区基因的一部分进行比较,证实了克隆的ε cDNA的身份。小鼠和大鼠的核苷酸序列有79%的同源性。

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