Abstract: Prokaryotic Expression and Purification of IL-13Rα2 Antigen in Human GliomasWenqiao YU1, Zuobing CHEN1, Yun ZHANG1, Zhongyan LIANG1, Shaoyang ZHANG1,Weiming ZHENG2Correspondence to:Weiming ZHENG, E-mail: zhwm61@126.com1Department of Emergency Surgery, The First Affiliated Hospital of Zhejiang University Medical School, Hangzhou 310000, China2Department of Neurosurgery, The First Affiliated Hospital ofWenzhou Medical College,Wenzhou 325000, ChinaAbstract Objective: To construct the expression plasmid of the Interleukin-13α2 Receptor (IL-13Rα2) and to detect in vitroprokaryotic expression of the receptor gene, in order to invetigate the theoretical basis of of immunotherapy for human gliomas.Methods: The object gene IL-13Rα2 was obtained from U251 glioma cell line by RT-PCR, then the cloning plasmid pMD19 Simple Tvector was connected and transfered to DH5α. Positive colonies were screened and recombinated. The IL-13Rα2 was digested fromrecombinant cloning plasmid using Xho I and Hind III, and was connected to the expression plasmid pET-28a that was also digested bythe same enzymes. It was then transfered to the BL21(DE3). After the recombinant plasmid was abstracted for digestion andsequencing, the prokaryotic expression of IL-13Rα2 antigenic peptide was conducted by the induction of IPTG. Nickel column wasused for recovery and purification, and SDS-PAGE was conducted to detect the protein expression. Results: The IL-13Rα2 antigen wassuccessfully expressed and the molecular weight was 60kD, which existed with the modality of inclusion bodies. Conclusion: Theobject gene IL-13Rα2 could be obtained from U251 glioma cell line by RT-PCR and connected to the expression plasmid pET-28a verysuccessfully. The expression plasmid of IL-13Rα2 gene can be successfully constructed in vitro and can be successfully expressed andpurified by IPTG induction to obtain the IL-13Rα2 antigenic peptide.Keywords Gliomas; IL-13Rα2 antigen; Expression plasmid