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从小鼠胚胎基因文库中克隆的免疫球蛋白γ2b重链基因的结构

Structure of immunoglobulin gamma 2b heavy chain gene cloned from mouse embryo gene library.

作者信息

Zakut R, Givol D, Mory Y Y

出版信息

Nucleic Acids Res. 1980 Feb 11;8(3):453-66. doi: 10.1093/nar/8.3.453.

Abstract

A mouse DNA clone containing the constant part of the immunoglobulin gamma 2b heavy chain was isolated from a mouse gene library. The library was constructed in Charon 4A from a partial EcoRI digest of mouse embryo DNA and was screened with a plasmid (p gamma (11)7) containing a cDNA insert of the heavy chain constant region of the plasmacytoma MPC-11 (1). The Charon 4A clone contains a 14 kb insert which is cleaved by EcoRI into a 6.8 kb and 7.2 kb fragments, of which only the 6.8 kb contains the sequence for gamma 2b heavy chain. Restriction analysis and partial sequence of the insert in p gamma (11) 7 enabled us to obtain three fragments corresponding to the 5' (amino acid 161-302) middle (amino acid 302-443) and 3' (mostly non coding 107 bp) regions of the constant region. Restriction analysis of the Charon 4A clone and hybridisation to these nick translated fragments revealed that the gamma 2b constant region gene contains about 1.5 kb and has three intervening sequences.

摘要

从一个小鼠基因文库中分离出一个包含免疫球蛋白γ2b重链恒定区的小鼠DNA克隆。该文库是用来自小鼠胚胎DNA的部分EcoRI酶切片段构建于Charon 4A载体上,并使用含有浆细胞瘤MPC - 11重链恒定区cDNA插入片段的质粒(pγ(11)7)进行筛选的。Charon 4A克隆含有一个14 kb的插入片段,该片段被EcoRI酶切成一个6.8 kb和一个7.2 kb的片段,其中只有6.8 kb的片段包含γ2b重链的序列。对pγ(11)7中插入片段的限制性分析和部分测序使我们获得了与恒定区5'(氨基酸161 - 302)、中间(氨基酸302 - 443)和3'(大部分为非编码的107 bp)区域相对应的三个片段。对Charon 4A克隆的限制性分析以及与这些缺口平移片段的杂交显示,γ2b恒定区基因约为1.5 kb,并有三个间隔序列。

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