title: [Cloning and expression of human interleukin-26 in Escherichia coli].
abstract: To clone human interleukin-26 (hIL-26) and express it in E. coli efficiently. Two pairs of primers were synthesized according to the hIL-26 gene reported on GenBank. The hIL-26 gene was cloned by nest PCR following the first round RT-PCR from human peripherial blood monocytes total RNA, and then the PCR product was cloned into pMD18-T vector. Colony PCR, restriction analysis and sequence analysis showed that the gene cloned was the same as the reported hIL-26. The recombinant was cut with BamHI and EcoR I to obtain the hIL-26 fragment, and then the fragment was inserted into pBV220 which was cut with the same enzymes. The recombinant expression vector was induced to express hIL-26 at 42 degrees C, SDS-PAGE analysis showed that the recombinant protein accounted for up to 20% of the whole protein of E. coli, and the protein was also confirmed by Western blotting. Purity of the protein was found to be above 90% after purified with molecular sieve. After renaturalized with glutathione buffer, the promoting effect of it on the production of IFN-y in PBMC was detected by RT-PCR. A recombinant bacterial strain for expressing hIL-26 with biological activity was constructed successfully.
authors.affiliation: Medical Research Center, Shandong Provincial Hospital, Shandong University, Jinan 250021, China.
authors.forename: Yi-Qing
authors.initials: YQ
authors.lastname: Liu
authors.forename: Zi-Jiang
authors.initials: ZJ
authors.lastname: Chen
authors.forename: Xue
authors.initials: X
authors.lastname: Zhang
authors.forename: Lai-Cheng
authors.initials: LC
authors.lastname: Wang
authors.forename: Yu-Lian
authors.initials: YL
authors.lastname: Jiao
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.forename: Chun-Yan
authors.initials: CY
authors.lastname: Ma
authors.forename: Bin
authors.initials: B
authors.lastname: Cui
authors.forename: Xin-Pu
authors.initials: XP
authors.lastname: Gao
authors.forename: Zheng-Min
authors.initials: ZM
authors.lastname: Liu
authors.forename: Kan
authors.initials: K
authors.lastname: Wu
authors.forename: Yue-Ran
authors.initials: YR
authors.lastname: Zhao
mesh_terms: D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007378:Interleukins / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: C406767:IL26 protein, human; D007378:Interleukins; D011994:Recombinant Proteins