| 摘要 |
<p>According to the known cDNA sequence we used the PCR (polymerase chain reaction) technique to isolate a gene encoding human MIF from uterus endometrium The PCR product was cloned in the Escherichia coli plasmid vector pUC 19 and the nucleotide sequence was confirmed. We were able to successfully express human MIF in the bacteria Escherichia coli by use of the pKP 1500 expression plasmid. The recombinant protein accumulated intracellularly in soluble form, comprising more then 30 % of total cell protein. We have designed an original two step procedure where protein purification was accomplished by gel filtration and ion exchange chromatography. The purified recombinant MIF was used to immunise a rabbit and antibodies obtained were used for MIF detection in human tissues by immunohistochemical techniques.</p> |
