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Cloning of Rat TARC cDNA and Analysis of Tissue-Specific mRNA Expression

机译:大鼠TARC cDNA的克隆及组织特异性mRNA表达分析

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摘要

Thymus- and activation-regulated chemokine (TARC) is one that selectively controls the migration of type 2-helper T lymphocytes into inflammatory lesions. TARC is a CC chemokine and plays an essential role in recruiting CC chemokine receptor 4-positive Th2 cells to allergic lesions. We cloned TARC cDNA from rat thymus using RT-PCR. The rat TARC clone contained a full-length open reading frame encoding 93 amino acids that showed 83 and 66% homology with mouse and human homologs, respectively. The expression of TARC mRNA was mainly in the lymphoid organs, for example, the thymus, spleen, and lymph node. The recombinant TARC was expressed in Escherichia coli and purified in an active form. In addition, the purified rat TARC with S-tagged specifically binds to human CCR4 in CD4/CCR4-transfected HOS cells by cell-binding assay using flow cytometry. The TARC cDNA clones obtained in this study will be valuable for future studies on allergic diseases in rats.
机译:胸腺和激活调节趋化因子(TARC)是一种选择性控制2型辅助T淋巴细胞向炎症性病变迁移的趋化因子。 TARC是一种CC趋化因子,在将CC趋化因子受体4阳性Th2细胞募集到过敏性病变中起重要作用。我们使用RT-PCR从大鼠胸腺克隆了TARC cDNA。大鼠TARC克隆包含一个全长开放阅读框,该阅读框编码93个氨基酸,分别与小鼠和人类同源物有83%和66%的同源性。 TARC mRNA的表达主要在淋巴器官,例如胸腺,脾脏和淋巴结中。重组TARC在大肠杆菌中表达并以活性形式纯化。另外,通过使用流式细胞术的细胞结合测定,具有S-标记的纯化的大鼠TARC与CD4 / CCR4转染的HOS细胞中的人CCR4特异性结合。在这项研究中获得的TARC cDNA克隆将对大鼠变态反应性疾病的未来研究具有宝贵的价值。

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