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KMID : 0880220140520070604
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Journal of Microbiology
2014 Volume.52 No. 7 p.604 ~ p.608
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An easy way for the rapid purification of recombinant proteins from Helicobacter pylori using a newly designed expression vector
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Kang Hyung-Lyun
Jo Jin-Sung
Kwon Soon-Uck
Song Jae-Young
Seo Ji-Hyun
Cho Myung-Je
Baik Seung-Chul
Youn Hee-Shang
Rhee Kwang-Ho
Lee Woo-Kon
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Abstract
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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 pBKcontained 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 themulti-cloning site of pBK. The orf of cat was inserted downstreamof 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.
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KEYWORD
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Helicobacter pylori, pBKHGC, vector
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