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来自B细胞和小鼠-人杂交瘤的人免疫球蛋白μ链cDNA的克隆及部分核苷酸序列

Cloning and partial nucleotide sequence of human immunoglobulin mu chain cDNA from B cells and mouse-human hybridomas.

作者信息

Dolby T W, Devuono J, Croce C M

出版信息

Proc Natl Acad Sci U S A. 1980 Oct;77(10):6027-31. doi: 10.1073/pnas.77.10.6027.

Abstract

Purified mRNAs coding for mu and kappa human immunoglobulin polypeptides were translated in vitro and their products were characterized. The mu-specific mRNAs, derived from both human lymphoblastoid cells (GM607) and from a mouse-human somatic cell hybrid secreting human mu chains (alpha D5-H11-BC11), were copied into cDNAs and inserted into the plasmid pBR322. Several recombinant cDNAs that were obtained were identified by a combination of colony hybridization with labeled probes, in vitro translation of plasmid-selected mu mRNAs, and DNA nucleotide sequence determination. One recombinant DNA, for which the sequence has been partially determined, contains the codons for part of the C3 constant region domain through the carboxy-terminal piece (155 amino acids total) as well as the entire 3' noncoding sequence up to the poly(A) site of the human mu mRNA. The sequence A-A-U-A-A occurs 12 nucleotides prior to the poly(A) addition site in the human mu mRNA. Considerable sequence homology is observed in the mouse and human mu mRNA 3' coding and noncoding sequences.

摘要

编码人μ和κ免疫球蛋白多肽的纯化mRNA在体外进行翻译,并对其产物进行了表征。来自人淋巴母细胞(GM607)和分泌人μ链的小鼠-人体细胞杂种(αD5-H11-BC11)的μ特异性mRNA被复制成cDNA,并插入质粒pBR322中。通过用标记探针进行菌落杂交、质粒选择的μmRNA的体外翻译以及DNA核苷酸序列测定相结合的方法,鉴定了获得的几种重组cDNA。其中一个重组DNA的序列已部分确定,它包含从C3恒定区结构域的一部分到羧基末端片段(总共155个氨基酸)的密码子,以及直至人μmRNA的聚腺苷酸位点的整个3'非编码序列。序列A-A-U-A-A出现在人μmRNA中聚腺苷酸添加位点之前12个核苷酸处。在小鼠和人μmRNA的3'编码和非编码序列中观察到相当程度的序列同源性。

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