多发性骨髓瘤细胞中一个表达上调基因的克隆与分析

Molecular Cloning and Analysis of a Novel Gene Over-expressed in Multiple Myeloma Cells

根据GenBank收录的多发性骨髓瘤细胞株 (ARH 77)表达上调ESTAF4 2 5 30 0设计引物 ,运用RT PCR检测了 5例多发性骨髓瘤患者及 4例正常人骨髓细胞中该EST的表达水平 .Northern印迹杂交分析该EST在多种组织中的表达 .进一步利用该EST作探针 ,筛选ARH 77cDNA文库 ,获得全长cDNA克隆 ,对该序列进行了分析 .结果显示 ,该EST在多发性骨髓瘤患者骨髓细胞中亦有较高的表达 ,而在正常人骨髓细胞中低表达 .经测序证实 ,该cDNA全长为 4 5 2bp(GenBank收录号 :AF4 87338) .预测其编码一个 5 7个氨基酸的小分子量蛋白质 ,属于与DNA复制有关的解旋酶 引物酶基因家族的新成员 .该基因在多发性骨髓瘤细胞中表达上调 。

According to the over expressed EST AF425300, a pair of primers were designed for detecting the expression level of EST AF425300 in multiple myeloma(MM) bone marrow cells from 5 patiens and 4 normal bone marrow samples as control by using RT PCR. AF425300 was over expressed in MM samples and expressed on a low level in control. The expression of AF425300 in 12 kinds of human tissues was also determined by Northern blot analysis. The full length cDNA clone was screened out from ARH 77 cDNA library and sequenced. The results were analyzed by bioinformatics. A protein of 57 amino acid residues was deduced from the 452 bp full length cDNA and may be a novel member of helicase/primase family. It is overexpressed in MM cells and might be involved in the development of MM.