An Easy Way for the Rapid Purification of Recombinant Proteins from Helicobacter pylori Using a Newly Designed Expression Vector

Hyung-Lyun Kang; Jin-Sung Jo; Soon-Uck Kwon; Jae-Young Song; Ji-Hyun Seo; Myung-Je Cho; Seung-Chul Baik; Hee-Shang Youn; Kwang-Ho Rhee; Woo-Kon Lee

doi:10.1007/s12275-014-3679-y

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Research Support, Non-U.S. Gov't
An Easy Way for the Rapid Purification of Recombinant Proteins from Helicobacter pylori Using a Newly Designed Expression Vector: Hyung-Lyun Kang ^1,2, Jin-Sung Jo ³, Soon-Uck Kwon ¹, Jae-Young Song ^1,2, Ji-Hyun Seo ⁴, Myung-Je Cho ^1,2, Seung-Chul Baik ^1,2, Hee-Shang Youn ⁴, Kwang-Ho Rhee ^1,2, Woo-Kon Lee ^1,2; Journal of Microbiology 2014;52(7):604-608
DOI: https://doi.org/10.1007/s12275-014-3679-y
Published online: June 28, 2014

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 Korea¹Department 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

Corresponding author: Woo-Kon Lee , Tel: +82-55-772-8083,

Received: 26 December 2013 • Revised: 8 May 2014 • Accepted: 15 May 2014

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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.

Keywords: Helicobacter pylori; pBKHGC; vector

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An Easy Way for the Rapid Purification of Recombinant Proteins from Helicobacter pylori Using a Newly Designed Expression Vector

J. Microbiol. 2014;52(7):604-608. Published online June 28, 2014

DOI: https://doi.org/10.1007/s12275-014-3679-y

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