Journal: PLoS ONE
Article Title: Protection against Th17 Cells Differentiation by an Interleukin-23 Receptor Cytokine-Binding Homology Region
Figure Lengend Snippet: Generation of IL23R-CHR protein. A, Restriction sites are indicated in bold, the translated amino acid and Trx-6×His-DDDDKA tag are shown in the box; B, IL23R-CHR gene from human spleen cDNA library amplified by PCR; C, One pET32a/IL23R-CHR clone digested with NcoI and XhoI and analyzed by agrose gel electrophoresis (1%). MW = molecular weight markers, bp. D, Trx-IL23R-CHR induced by IPTG in E.Coli BL21 (DE3), SDS-PAGE (15%), protein bands were stained with coomassie brilliant blue R250 reagent. Lane 2, uninduced bacterial lysate; Lane 3, IPTG induced bacterial lysate. E, Trx-IL23R-CHR purified from BL21 (DE3) lysate and cleaved by enterokinase, Lane 5, purified Trx-IL23R-CHR; Lane 6, purified IL23R-CHR. F, Western blot using mouse mAbs against human IL23R (Lane 8). Lane 1, 4 and 7 in D, E, F: molecular weight markers, KD.
Article Snippet: After purification with Tianquick midi purification kit (Tiangen, China), the IL23R-CHR gene was digested with NcoI and XhoI (takara) and inserted into pET32a following the manufacturer’s instructions (Novagen, Germany).
Techniques: cDNA Library Assay, Amplification, Polymerase Chain Reaction, Nucleic Acid Electrophoresis, Molecular Weight, SDS Page, Staining, Purification, Western Blot