| Title | An Easy Way for the Rapid Purification of Recombinant Proteins from Helicobacter pylori Using a Newly Designed Expression Vector |
|---|---|
| Author | Hyung-Lyun Kang1,2, Jin-Sung Jo3, Soon-Uck Kwon1, Jae-Young Song1,2, Ji-Hyun Seo4, Myung-Je Cho1,2, Seung-Chul Baik1,2, Hee-Shang Youn4, Kwang-Ho Rhee1,2, and Woo-Kon Lee1,2* |
| Address | 1Department of Microbiology, Gyeongsang National University School of Medicine, Jinju 660-751, Republic of Korea, 2Research Institute of Life Science, Gyeongsang National University, Jinju 660-701, Republic of Korea, 3Korea Forest Research Institute, Suwon, 441-350, Republic of Korea, 4Department of Pediatrics, Gyeongsang National University School of Medicine, Jinju 660-751, Republic of Korea |
| Bibliography | Journal of Microbiology, 52(7),604-608, 2014, |
| DOI | 10.1007/s12275-014-3679-y |
| Key Words | Helicobacter pylori, pBKHGC, vector |
| Abstract | We constructed a H. pylori expression vector which consisted of both a His-tag and a GST tag as purification tools for recombinant protein and a chloramphenicol resistant cat gene as a reporter. The backbone of the vector pBK contained an ColEI origin of replication and a kanamycin resistant gene. A set of oligos for the His-tag and the PCR product of gst (glutathione S-transferase) gene were inserted sequentially in frame in the multi-cloning site of pBK. The orf of cat was inserted downstream of the gst to generate pBKHGC. The 3' part of H. pylori clpB and flaA were cloned into the vector which was introduced into H. pylori. Recombinant proteins were purified by GSH affinity column, digested with thrombin and were analyzed by western blotting. The final recombinant proteins were successfully purified. |