2001•Europe PMC (PubMed Central)Requires access
Construction of recombinant human interleukin 18 expression vector and its high level expression in E. coli
Ruijun Zhang, Tian Zhigang, Wei Haiming, Rui Sun, Jinbo Feng, Yueran Zhao
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Abstract
PurposeThe aim is to clone human IL 18 cDNA,to construct recombinant human IL 18 expression vector and to perform the bioassay of recombinant human IL 18 (rhIL 18) expressed in E.coli. Methods Human IL 18 cDNA fragment (496 bp) was amplified from PBMC by RT PCR method and sequenced by ABI PRIM TM 377XL DNA Sequencer. The cloned IL 18 cDNA fragment was inserted into pBV220 expression vector, which was then transformed to E.coli. SDS PAGE and Western blot assays showed that rhIL 18 was highly expressed. The bioassay on rhIL 18 was performed by MTT assay in vitro. ResultsThe expression level of recombinant IL 18 protein reached 27% and the purity is over 96% after purification. The refolded rhIL 18 enhanced the cytotoxicity of NK cells in the presence of IL 2(10 U/ml) in vitro. ConclusionThe expressed rhIL 18 is functional and can be applied for further studies.