Purification and characterization of recombinant human interleukin-29 expressed in Escherichia coli.
Human interleukin (IL)-29 is the latest member of the class II cytokine family. However, as a result of lacking efficient method to generate relatively large quantity of IL-29, little is known of its functions in man. In the present study, an Escherichia coli expression system for the rapid expression of the human IL-29 gene was developed. It involved of cloning IL-29 gene into the pET-44 Ek/LIC vector, which allowed expression of IL-29 with a fusion tag consisting of the NusA protein, polyhistidine and S peptide (Nus-His-S-tag), and introducing a thrombin recognition site between the fusion tag and IL-29. The expressed fusion protein was purified by S-protein agarose affinity chromatography, and the fusion tag was removed from recombinant IL-29 by cleavage with thrombin. The purified IL-29 appeared a single band on SDS-PAGE, and the yield of IL-29 was 60 mg from 1 l of bacterial culture. N-terminal sequencing confirmed the identity of the purified protein. The recombinant IL-29 showed specific antiviral activity that was comparable to the commercially available IFN alfa-2b preparation.
Li
Mingcai
M
Allergy and Inflammation Research Institute, Shantou University Medical College, 22 Xin-Ling Road, Shantou 515041, Guangdong Province, People's Republic of China.
He
Shaoheng
S
D002460:Cell Line; D016207:Cytokines / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors; D006801:Humans; D007372:Interferons; D007378:Interleukins / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D013917:Thrombin / Q000737:chemistry; D014721:Vesicular stomatitis Indiana virus / Q000187:drug effects / Q000472:pathogenicity; D000095022:Interferon Lambda
D016207:Cytokines; D007372:Interferons; D007378:Interleukins; D011993:Recombinant Fusion Proteins; D013917:Thrombin; C000624353:interferon-lambda, human; D000095022:Interferon Lambda