2007•Biotechnology(Faisalabad)Requires access
Cloning and Prokaryotic Expression of Human Interleukin-10
Wang XingLong
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Abstract
Objective:To construct Prokaryotic expression vector of human IL-10 and identify the expressed production.Methods:a cDNA encoding human interleukin-10(IL-10) was amplified by RT-PCR from lymphocyte actived by ConA.the IL-10 cDNA was cloned to vector PGEM-T.After confirming the sequence,IL-10 cDNA was extracted and construct the Prokaryotic expression vector(PET28a-IL10).The E.coli BL21(DE3) was transfected with the constructed PET28a-IL10,and the expressed IL-10 protein was assayed by SDS-PAGE and Western-blot.Results:SDS-PAGE and Westernblot show that the IL-10 protein,with a relative molecular weight of 21000,was successfully expressed and can specifically combine with the rabbit anti-human IL-10 antibody.Conclusion:IL-10 protein was expressed in E.coli and showed good specificity.which laid a foundation of IL-10 further biololgy rsearch.