MOLECULAR-CLONING OF A CDNA-ENCODING A HUMAN MACROPHAGE-MIGRATION INHIBITORY FACTOR
MOLECULAR-CLONING OF A CDNA-ENCODING A HUMAN MACROPHAGE-MIGRATION INHIBITORY FACTOR
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DOI：
10.1073/pnas.86.19.7522
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发表时间：
1989-10-01
影响因子：
11.1
通讯作者：
DAVID, JR
中科院分区：
文献类型：
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作者：
WEISER, WY;TEMPLE, PA;DAVID, JR
通过在COS-1细胞中的功能性表达克隆，从由凝集素刺激的T细胞杂交瘤T-CEMB制备的cDNA文库中分离编码人巨噬细胞移动抑制因子（MIF）的cDNA。由MIF cDNA（p7-1）编码的115个氨基酸的多肽从转染的COS-1细胞中有效地释放，并且在培养上清液中产生容易检测的MIF活性，尽管明显缺乏经典的蛋白质分泌序列。插入突变分析和从聚丙烯酰胺凝胶切片中洗脱MIF活性证明COS-1细胞释放的具有MIF活性的Mr 12，000蛋白由p7-1编码。p7-1 cDNA与Con-A刺激的淋巴细胞表达的700个碱基的mRNA杂交，但不与未刺激的淋巴细胞杂交。MIF cDNA克隆和重组MIF的可用性将有助于分析这种淋巴因子在细胞介导的免疫、免疫调节和炎症中的作用。
A cDNA encoding a human macrophage migration inhibitory factor (MIF) was isolated, through functional expression cloning in COS-1 cells, from a cDNA library prepared from a lectin-stimulated T-cell hybridoma, T-CEMB. The 115-amino acid polypeptide encoded by the MIF cDNA (p7-1) was effectively released from the transfected COS-1 cells and yielded readily detectable MIF activity in the culture supernatant despite the apparent lack of a classical protein secretory sequence. Insertional mutational analysis and elution of MIF activity from polyacrylamide gel slices demonstrated that the Mr 12,000 protein with MIF activity released by the COS-1 cells is encoded by p7-1. The p7-1 cDNA hybridized with a 700-base mRNA expressed by Con-A-stimulated lymphocytes but not unstimulated lymphocytes. The availability of the MIF cDNA clone and recombinant MIF will facilitate the analysis of the role of this lymphokine in cell-mediated immunity, immunoregulation, and inflammation.