Journal of Microbiology

Kwang-Ho Rhee

5 Articles

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; 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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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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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

Profiling of the Bacteria Responsible for Pyogenic Liver Abscess by 16S rRNA Gene Pyrosequencing: Yun Gyu Song , Sang Gun Shim , Kwang Min Kim , Dong-Hae Lee , Dae-Soo Kim , Sang-Haeng Choi , Jae-Young Song , Hyung-Lyun Kang , Seung-Chul Baik , Woo-Kon Lee , Myung-Je Cho , Kwang-Ho Rhee; J. Microbiol. 2014;52(6):504-509. Published online May 29, 2014; DOI: https://doi.org/10.1007/s12275-014-4241-7

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Abstract PDF

Pyogenic liver abscess (PLA) is a severe disease with considerable mortality and is often polymicrobial. Understanding the pathogens that cause PLA is the basis for PLA treatment. Here, we profiled the bacterial composition in PLA fluid by pyrosequencing the 16S ribosomal RNA (rRNA) gene based on next-generation sequencing (NGS) technology to identify etiological agents of PLA and to provide information of their 16S rRNA sequences for application to DNA-based techniques in the hospital. Twenty patients with PLA who underwent percutaneous catheter drainage, abscess culture, and blood culture for isolates were included. Genomic DNAs from abscess fluids were subjected to polymerase chain reaction and pyrosequencing of the 16S rRNA gene with a 454 GS Junior System. The abscess and blood cultures were positive in nine (45%) and four (20%) patients, respectively. Pyrosequencing of 16S rRNA gene showed that 90% of the PLA fluid samples contained single or multiple genera of known bacteria such as Klebsiella, Fusobacterium, Streptococcus, Bacteroides, Prevotella, Peptostreptococcus, unassigned Enterobacteriaceae, and Dialister. Klebsiella was predominantly found in the PLA fluid samples. All samples that carried unassigned bacteria had 26.8% reads on average. We demonstrated that the occurrence of PLA was associated with eight known bacterial genera as well as unassigned bacteria and that 16S rRNA gene sequencing was more useful than conventional culture methods for accurate identification of bacterial pathogens from PLA.

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Genetic Organization and Conjugal Plasmid DNA Transfer of pHP69, a Plasmid from a Korean Isolate of Helicobacter pylori: 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; J. Microbiol. 2013;51(6):889-889.; DOI: https://doi.org/10.1007/s12275-013-0727-y

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Abstract PDF: 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: 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; J. Microbiol. 2012;50(6):955-961. Published online December 30, 2012; DOI: https://doi.org/10.1007/s12275-012-2580-9

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Abstract PDF

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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Plasmid.2019; 106: 102442. CrossRef
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Esther Fernandez-Gonzalez, Steffen Backert
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Helicobacter pylori γ-Glutamyltranspeptidase Induces Cell Cycle Arrest at the G1-S Phase Transition: Kyung-Mi Kim , Seung-Gyu Lee , Jung-Min Kim , Do-Su Kim , Jea-Young Song , Hyung-Lyun Kang , Woo-Kon Lee , Myung-Je Cho , Kwang-Ho Rhee , Hee-Shang Youn , Seung-Chul Baik; J. Microbiol. 2010;48(3):372-377. Published online June 23, 2010; DOI: https://doi.org/10.1007/s12275-010-9293-8

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Abstract PDF

In our previous study, we showed that Helicobacter pylori γ-glutamyltranspeptidase (GGT) is associated with H. pylori-induced apoptosis through a mitochondrial pathway. To better understand the role of GGT in apoptosis, we examined the effect of GGT on cell cycle regulation in AGS cells. To determine the effect of recombinant GGT (rGGT) on cell cycle distribution and apoptosis, rGGT-treated and untreated AGS cells were analyzed in parallel by flow cytometry using propidium iodide (PI). We found that rGGT inhibited the growth of AGS cells in a time-dependent manner, and that the pre-exposure of cells to a caspase-3 inhibitor (z-DEVD-fmk) effectively blocked GGT-induced apoptosis. Cell cycle analysis showed G1 phase arrest and apoptosis in AGS cells following rGGT treatment. The rGGT-mediated G1 phase arrest was found to be associated with down-regulation of cyclin E, cyclin A, Cdk 4, and Cdk 6, and the up-regulation of the cyclindependent kinase (Cdk) inhibitors p27 and p21. Our results suggest that H. pylori GGT induces cell cycle arrest at the G1-S phase transition.

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