[Construction of recombinant expression vector of IL-24 gene and prokaryotic expression].
OBJECTIVE
To construct a recombinant expression vector of human IL-24 gene and express it in E. coli.


METHODS
The hIL-24 cDNA fragment was amplified from plasmid TRAP-hIL-24 by PCR, then cloned into the prokaryotic vector pGEX-KG, and expressed as a fusion protein in E. coli. The expressed IL-24-GST fusion protein was purified via GST-Sepharose 4B Column and identified by SDS-PAGE and Western blot. The bioactivity of GST-IL-24 fusion protein was measured by MTT assay.


RESULTS
Restriction enzyme digestion analysis showed that the recombinant prokaryotic expression vector pGEX-KG-IL-24 was successfully constructed and expressed in E. coli. The relative molecular mass (M(r)) of the expression product was identical with the predicted value. The proliferation of THP-1 cells was inhibited by GST-IL-24 fusion protein.


CONCLUSIONS
The recombinant expression vector pGEX-KG-IL-24 has been constructed successfully and expressed as a bioactive fusion protein in E. coli BL21 (BlysS), which is helpful for the further study of the biological function of IL-24.
Qu
Shao-gang
SG
Department of Immunology, Medical College of Wuhan University, Wuhan 430071, China.
Chen
Ming
M
Zhang
Xiao-lian
XL
D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D007378:Interleukins / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D011387:Prokaryotic Cells / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D000099180:Interleukin-24
D007378:Interleukins; D011993:Recombinant Fusion Proteins; D000099180:Interleukin-24