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为了克隆新的细胞因子,根据白细胞介素-10(IL-10)能够广谱抑制细胞因子表达和U937细胞在PHA刺激下能够产生多种细胞因子的原理,利用抑制性递减杂交(SSH)技术,研究了U937细胞中受IL-10抑制表达的新基因,发现了6个新的EST片段,对其中1个以后被证实的新CC家族趋化因子的序列通过EST拼接及RT-PCR扩增,得到了全长cD.NA.Northem Blot表明该分子大小与预期的一致,其表达可被IL-10所抑制,将其命名为UCK-1(U937细胞来源的趋化因子-1).UCK-1全长cDNA为534个碱基,包括polyA和AT-
In order to clone new cytokines, the principle that interleukin-10 (IL-10) can inhibit the expression of cytokines broadly and that U937 cells are capable of producing various cytokines under the stimulation of PHA, the use of suppression subtractive hybridization (SSH) , A novel gene that is inhibited by IL-10 in U937 cells was studied and 6 new ESTs were found. One of these newly identified CC family chemokines was amplified by EST splicing and RT-PCR , Giving the full-length cD.NA.Northem Blot showed that the size of the molecule was as expected and its expression was inhibited by IL-10 and named UCK-1 (U937 cell-derived chemokine-1) -1 The full-length cDNA is 534 bases, including polyA and AT-