In order to study the property of recombinant human macrophage migration inhibitory factor(MIF), we cloned MIF cDNA and expressed it using pET expression system.
MIF cDNA, about 380 bp, was amplified from total RNA of peripheral blood mononuclear cell by primers and reverse transcription polymerase chain reaction. cDNA was cloned to t vector and pET 3a plasmid and its sequences was determined by using
dideoxy
chain termination method. Homology search was done using gene bank and its sequence was shown to be identical to the human MIF cDNA sequence reported previously. MIF cDAN cloned to pET plasmid was introduced into BL21(DE30 competent cell, and its
expression was induced by adding IPTG. Production of recombinant MIF, about 14kD soluble protein, was gradually increased about 30 minutes after induction.
The recombinant MIF, anti-MIF antibody, MIF primers and MIF cDNA can be used in such many studies as search of new MIF function, estimation of MIF gene expression, identification of MIF synthesizing cells and cloning of MIF receptor gene in
future.
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