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[克隆编码抗人白细胞介素-2单克隆抗体轻链和重链可变区的cDNA片段]

[Cloning fragments of cDNA coding variable segments of light and heavy chains of monoclonal antibodies against human interleukin-2].

作者信息

Paskhin A I, Golovina T N, Nesmeianov V A, Korobko V G

出版信息

Bioorg Khim. 1995 Jun;21(6):430-5.

PMID:7661867
Abstract

DNA fragments coding for the variable fragments of the heavy and light chains of the immunoglobulin G1 against human recombinant interleukin-2 were produced using reverse transcription of the total RNA isolated from the murine hybrid myeloma cell line LNKB-2 followed by amplification of the RNA-DNA duplexes with degenerated primers. The fragments obtained were cloned into the plasmid pGEM7-Zf(+) and their structures were determined. The fragments cloned were proved to encode the Fv-fragments by sequencing N-termini of the light and heavy chains of the antibody.

摘要

通过逆转录从鼠杂交骨髓瘤细胞系LNKB - 2中分离的总RNA,随后用简并引物扩增RNA - DNA双链体,产生了编码针对人重组白细胞介素 - 2的免疫球蛋白G1重链和轻链可变片段的DNA片段。将获得的片段克隆到质粒pGEM7 - Zf(+)中,并确定其结构。通过对抗体轻链和重链的N端进行测序,证明克隆的片段编码Fv片段。

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