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利用聚合酶链反应克隆和测序人λ免疫球蛋白基因

Cloning and sequencing of human lambda immunoglobulin genes by the polymerase chain reaction.

作者信息

Songsivilai S, Bye J M, Marks J D, Hughes-Jones N C

机构信息

Molecular Immunopathology Unit, Medical Research Council Centre, Cambridge, GB.

出版信息

Eur J Immunol. 1990 Dec;20(12):2661-6. doi: 10.1002/eji.1830201220.

Abstract

Universal oligonucleotide primers, designed for amplifying and sequencing genes encoding the rearranged human lambda immunoglobulin variable region, were validated by amplification of the lambda light chain genes from four human heterohybridoma cell lines and in the generation of a cDNA library of human V lambda sequences from Epstein-Barr virus-transformed human peripheral blood lymphocytes. This technique allows rapid cloning and sequencing of human immunoglobulin genes, and has potential applications in the rescue of unstable human antibody-producing cell lines and in the production of human monoclonal antibodies.

摘要

设计用于扩增和测序编码重排的人λ免疫球蛋白可变区基因的通用寡核苷酸引物,通过从四个人源杂交瘤细胞系中扩增λ轻链基因以及从爱泼斯坦-巴尔病毒转化的人外周血淋巴细胞中生成人Vλ序列的cDNA文库进行了验证。该技术可实现人免疫球蛋白基因的快速克隆和测序,在拯救不稳定的人抗体产生细胞系以及生产人单克隆抗体方面具有潜在应用。

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