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小鼠胸腺细胞B细胞抗原(ThB)cDNA克隆的分离与鉴定

Isolation and characterization of cDNA clones for the mouse thymocyte B cell antigen (ThB).

作者信息

Gumley T P, McKenzie I F, Kozak C A, Sandrin M S

机构信息

Austin Research Institute, Austin Hospital, Heidelberg, Victoria, Australia.

出版信息

J Immunol. 1992 Oct 15;149(8):2615-8.

PMID:1401899
Abstract

The Thb locus is responsible for the expression of 15-kDa phosphatidyl inositol anchored molecules (ThB) on murine thymocytes and B cells. Thb expression as detected with mAb is polymorphic on B cells with two alleles, Thbh and Thb1 responsible for the high and low expression of ThB on B cells. The regulatory locus for Thb expression had been mapped with the Ly-6 cluster of genes to Chr 15. In our study we used expression cloning in COS cells to isolate cDNA clones that code for ThB after transfection; the cDNA products react with anti- ThB antibodies, but not with Ly-6A.2, -6B.2, -6C.2, or -6D.2 antibodies. One of these clones, pThB-A contains insert of 702 bases which was sequenced. The translated amino acid sequence has 11 cysteine residues, and together with the absence of potential N-linked glycosylation sites is similar to the structure of the Ly-6 molecules. The nucleotide and amino acid sequences of ThB cDNA were compared to those of Ly-6 genes and the Ly-6 related human CD59 and show clear homology. Finally using interspecies crosses, the structural Thb gene has been mapped to Chr 15; thus both structural and regulatory genes map to a similar site. The genetic map location near Ly-6 and the sequence similarity suggest that Thb and Ly-6 may have been derived from the same progenitor by gene duplication.

摘要

Thb基因座负责在小鼠胸腺细胞和B细胞上表达15-kDa磷脂酰肌醇锚定分子(ThB)。用单克隆抗体检测到的Thb表达在B细胞上具有多态性,有两个等位基因,Thbh和Thb1,分别负责B细胞上ThB的高表达和低表达。Thb表达的调控基因座已与Ly-6基因簇一起定位到15号染色体上。在我们的研究中,我们利用COS细胞中的表达克隆技术来分离转染后编码ThB的cDNA克隆;cDNA产物与抗ThB抗体反应,但不与Ly-6A.2、-6B.2、-6C.2或-6D.2抗体反应。其中一个克隆pThB-A包含一个702个碱基的插入片段,并进行了测序。翻译后的氨基酸序列有11个半胱氨酸残基,并且由于没有潜在的N-连接糖基化位点,其结构与Ly-6分子相似。将ThB cDNA的核苷酸和氨基酸序列与Ly-6基因以及与Ly-6相关的人类CD59的序列进行比较,显示出明显的同源性。最后,通过种间杂交,Thb结构基因已被定位到15号染色体上;因此,结构基因和调控基因都定位到了相似的位点。靠近Ly-6的遗传图谱位置和序列相似性表明,Thb和Ly-6可能是通过基因复制从同一个祖先进化而来的。

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