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功能性、抗原特异性抑制性T细胞杂交瘤中T细胞抗原受体α链基因转录本的序列与表达

Sequence and expression of transcripts of the T-cell antigen receptor alpha-chain gene in a functional, antigen-specific suppressor-T-cell hybridoma.

作者信息

Imai K, Kanno M, Kimoto H, Shigemoto K, Yamamoto S, Taniguchi M

出版信息

Proc Natl Acad Sci U S A. 1986 Nov;83(22):8708-12. doi: 10.1073/pnas.83.22.8708.

Abstract

We have detected rearrangement and expression of a gene encoding a T-cell antigen receptor alpha chain in the keyhole limpet hemocyanin (KLH)-specific, inducible suppressor-T-cell (Ts) hybridoma 34S-281 (BW5147 lymphoma-C57BL/6Ts) by using alpha-chain cDNA clones isolated from this Ts hybridoma. The cDNA clones have a restriction length polymorphism in the constant region that identifies them as being of C57BL/6 origin. The cDNA sequence has an ATG start codon for an open reading frame including variable, joining, and constant gene segments. Furthermore, the Ts alpha-chain gene transcripts were detected on membrane-bound polysomes by RNA blot analysis using a variable-region fragment from one of the alpha-chain cDNA clones as probe, suggesting that they are actively translated in the Ts hybridoma. As the beta-chain gene is deleted in all Ts hybridomas analyzed, and since a disulfide-linked dimer is detectable by two-dimensional NaDodSO4/PAGE of lysates of surface-radioiodinated Ts, we suggest that Ts antigen receptors are either alpha-chain homodimers or heterodimers composed of the alpha chain in association with an undefined chain.

摘要

我们通过使用从匙孔血蓝蛋白(KLH)特异性、可诱导抑制性T细胞(Ts)杂交瘤34S - 281(BW5147淋巴瘤 - C57BL/6Ts)中分离出的α链cDNA克隆,检测到了编码T细胞抗原受体α链的基因的重排和表达。这些cDNA克隆在恒定区具有限制性长度多态性,表明它们来源于C57BL/6。cDNA序列有一个用于开放阅读框的ATG起始密码子,该开放阅读框包括可变区、连接区和恒定基因片段。此外,使用α链cDNA克隆之一的可变区片段作为探针,通过RNA印迹分析在膜结合多核糖体上检测到了Tsα链基因转录本,这表明它们在Ts杂交瘤中被积极翻译。由于在所有分析的Ts杂交瘤中β链基因均缺失,并且通过表面放射性碘化Ts裂解物的二维NaDodSO4/PAGE可检测到二硫键连接的二聚体,我们推测Ts抗原受体要么是α链同二聚体,要么是由α链与一条未确定的链组成的异二聚体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8818/387000/281acfb37ad6/pnas00326-0282-a.jpg

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