- An Easy Way for the Rapid Purification of Recombinant Proteins from Helicobacter pylori Using a Newly Designed Expression Vector
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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
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J. Microbiol. 2014;52(7):604-608. Published online June 28, 2014
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DOI: https://doi.org/10.1007/s12275-014-3679-y
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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 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.
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Citations
Citations to this article as recorded by  - Gene polymorphisms of pathogenic Helicobacter pylori in patients with different types of gastrointestinal diseases
Yu-Li Chen, Xiao-Qiang Mo, Gan-Rong Huang, Yan-Qiang Huang, Juan Xiao, Li-Juan Zhao, Hong-Yu Wei, Qian Liang
World Journal of Gastroenterology.2016; 22(44): 9718. CrossRef
- Genetic Organization and Conjugal Plasmid DNA Transfer of pHP69, a Plasmid from a Korean Isolate of Helicobacter pylori
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Jung-Soo Joo , Jae-Young Song , Seung-Chul Baik , Woo-Kon Lee , Myung-Je Cho , Kon-Ho Lee , Hee-Shang Youn , Ji-Hyun Seo , Kwang-Ho Rhee , Hyung-Lyun Kang
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J. Microbiol. 2013;51(6):889-889.
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DOI: https://doi.org/10.1007/s12275-013-0727-y
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In the article by Joo et al. published in the Journal of Microbiology 2012; 50, 955-961. Dr. YoungAh Cho, one of coauthor’s
name was inadvertently missing. Complete authors information is as follow.
Jung-Soo Joo4, Jae-Young Song1,3, Seung-Chul Baik1,3, Woo-Kon Lee1,3, Myung-Je Cho1,3, Kon-Ho Lee1,3,
YoungAh Cho1, Hee-Shang Youn2, Ji-Hyun Seo2, Kwang-Ho Rhee1,3, and Hyung-Lyun Kang1,3*
- Genetic Organization and Conjugal Plasmid DNA Transfer of pHP69, a Plasmid from a Korean Isolate of Helicobacter pylori
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Jung-Soo Joo , Jae-Young Song , Seung-Chul Baik , Woo-Kon Lee , Myung-Je Cho , Kon-Ho Lee , Hee-Shang Youn , Ji-Hyun Seo , Kwang-Ho Rhee , Hyung-Lyun Kang
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J. Microbiol. 2012;50(6):955-961. Published online December 30, 2012
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DOI: https://doi.org/10.1007/s12275-012-2580-9
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We isolated pHP69, a 9,153 bp plasmid from Helicobacter pylori with a 33.98% (G+C) content. We identified 11 open reading frames (ORFs), including replication initiation protein A (repA), fic (cAMP-induced filamentation protein), mccC, mccB, mobA, mobD, mobB, and mobC, as well as four 22 bp tandem repeat sequences. The nucleic acid and predicted amino acid sequences of these ORFs exhibited significant homology to those of other H. pylori plasmids. pHP69 repA encodes a replication initiation protein and its amino acid sequence is similar to those of replicase proteins from theta-type plasmids. pHP69 contains two types of repeat
sequences (R1 and R2), a MOBHEN family mobilization region comprising mobC, mobA, mobB, and mobD, and genes encoding microcin B and C. Among the 36 H. pylori strains containing plasmids, mobA or mccBC are present in 12 or 6, respectively and 3 contain both genes. To examine intrinsic capability of H. pylori for conjugative plasmid transfer, a shuttle vector pBHP69KH containing pHP69 and replication origin of pBR322 was constructed. It was shown that this vector could stably replicate and be mobilized among clinical H. pylori strains and demonstrated to gene transfer by natural plasmid.
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Citations
Citations to this article as recorded by - Construction and characterization of the first Arcobacter butzleri - Escherichia coli shuttle vector
Adrián Salazar-Sánchez, Rodrigo Alonso, Aurora Fernández-Astorga, Ilargi Martínez-Ballesteros, Irati Martinez-Malaxetxebarria
Archives of Microbiology.2026;[Epub] CrossRef - Comprehensive plasmidomic analysis of Helicobacter pylori reveals the potential role of plasmids in pathogenic adaptation and a novel putative toxin-antitoxin system
Bradd Mendoza-Guido, Juan D Romero-Carpio, Silvia Molina-Castro
FEMS Microbiology Letters.2025;[Epub] CrossRef - Four Chromosomal Type IV Secretion Systems in Helicobacter pylori: Composition, Structure and Function
Wolfgang Fischer, Nicole Tegtmeyer, Kerstin Stingl, Steffen Backert
Frontiers in Microbiology.2020;[Epub] CrossRef - Two novel transcriptional reporter systems for monitoring Helicobacter pylori stress responses
A.M. Belova, D.V. Basmanov, V.V. Babenko, O.V. Podgorny, T.V. Mitko, K.A. Prusakov, D.V. Klinov, V.N. Lazarev
Plasmid.2019; 106: 102442. CrossRef - DNA transfer in the gastric pathogen Helicobacter pylori
Esther Fernandez-Gonzalez, Steffen Backert
Journal of Gastroenterology.2014; 49(4): 594. CrossRef
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