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이우곤,백승철,신민경,정명환,Jin-Sik Park,Jong-Hoon Ha,Dong-Hae Lee,Min Jeong Kim,Jeong-ih Shin,강형련 대한미생물학회 2019 Journal of Bacteriology and Virology Vol.49 No.4
In order to investigate the antioxidant effect of alkylhydroxide peroxidase (ahpC) of Helicobacter pylori (H. pylori) 26695, an ahpC-deficient mutant (H. pylori 26695 ahpC::cat) was generated. ahpC-deficient mutant was grown slowly at lower pressure of oxygen (5% oxygen) compared to the H. pylori 26695. Whole cell proteins isolated form H. pylori 26695 and H. pylori 26695 ahpC::cat were analyzed by MALDI-TOF and tandem-MS. The expression of 15 proteins, including Ppa, HypB, GrpE, Elp, RecA, GroES, Mda66, RibE, NapA, GlnA, BioB, TrxB, Tsf, FumC and Icd, was more than doubled in H. pylori 26695 ahpC::cat. Production of 10 proteins such as UreG, FabE, Adk, Pnp, OorC, AtpA, AtpD, Nqq3, Pfr, and TagD decreased below 50% in H. pylori 26695 ahpC::cat compared to the H. pylori 26695. In microarray analysis, 9 genes including sul1, amiE, frxA, fecA, hyuA, and katA increased in transcription level in H. pylori 26695 ahpC::cat compared to H. pylori 26695. A total of 24 genes, including flaB, protein kinase C inhibitor, cag16, pabC, and sabA, reduced in transcription. 27 genes, including HP0889, showed common expression changes in ahpC, katA, and sodB-deficient mutations. As a result of this study, there were not many genes whose expression was commonly changed by the deletion of each of the three major antioxidant enzymes of H. pylori. These results showed the functions and regulation of the three antioxidant enzymes were different in H. pylori.
Production of the Monoclonal Antibody and the Genomic Library of Helicobacter pylori
RHEE, KWANG HO,LEE, WOO KON 경상대학교 유전공학연구소 1990 遺傳工學硏究所報 Vol.9 No.-
Sixty three monoclonal antibodies(Moabs) to H. pylori were produced and classified into 13 groups according to their reaction pattern in Weatern blot analysis using whole cell lysate antigen of H. pylori. Genomic libary of H. pylori DNA was constructed to express recombinant antigen of H. pylori and 6 positive clons were obtained. Affinity-purified rabbit antibody to clone HRI and 2 reacted to 60 and 71kD antigen, respectively, and HR4 and 6 reacted to 66kD antigen in WEstern blot analysis. Antibody to HR3 reacted to several antigens, 71,66,60,55, and 34kD, and antibody to HR5 reacted to 71, 65, 60, 55, 49, and 34kD antigens. The one of MoAb, HPH216, reacted to HR4 recombinant antigen and 7 MoAbs, HPB3, 30, 33, 34, 36, 37 and 67, reacted to HR5 recombinant antigen. From the E. coli iysogens which have been lysogenized with 5 recombinant clones, 165kD fusion protein from HR4 and 180kD fusion protein with 71 and 29kD fragments from HR5 lysogen were detected.
Helicobacter pylori와 대장균의 Shuttle Vector 개발
조명제,이우곤,이상룡,김경희,안영숙,김성희,김현주,류복덕,최여정,윤영혜,백승철,전영석,이광호 경상대학교 유전공학연구소 1997 遺傳工學硏究所報 Vol.16 No.-
In this study, a vehicle vector using cryptic plasmids was constructed for gene transfer in Helicobacter pylori. pHP51(3.9 kb) and pHP489(1.2 kb) were selected for constructing vectors from cryptic plasmid of H. pylori isolates in Korea. The HindⅢ-digestedDNA fragment(1.2kb) of pHP489 and 1.6kb DNA fragment of pHP51 were ligated with a kanamycin resistance gene(aph3'-Ⅲ) from C. jejuni to produce the recombinant plasmids pHP489K and pHP51K, respectively. Transformation frequency of pHP51K by electroporation was low. But pHP489K could be effectively transformed into various H. pylori strains. In order to design an intermdiate vehicle vector for gene transfer into H. pylori, pBlueHP489K was prepared by recloning pHP489K DNA into pBluescript and pTZ19R vector. This vector permitted the DNA fragment containing pHP489 sequence, aph3'-Ⅲ, and cloned DNA to be cut and self-ligated in the SacⅠ site after cloning. ureA and ureB gene were inserted into pBlueHP489K, resulting in pBlueHP489K/AB. The DNA fragment containing pHP489, kanamycin resistance gene(aph3'-Ⅲ), and urease structural gene was cut away from pBlueHP489K/AB and self-ligated to generate pBlueHP489K/AB. pBlueHP489K/AB made urease-negative H. pylori strains restore their urease activity. By this experiment, pBlueHP489K was confirmed to be the vehicle system for transferring H. pylori genes.
Helicobacter pylori 편모 유전자의 클로닝 및 염기서열 분석
이광호,이우곤,조명제,도영미,백승철,강경희,박필성,이상룡 경상대학교 유전공학연구소 1993 遺傳工學硏究所報 Vol.12 No.-
A λgt11 expression libary of H. pylori DNA in E. coli Y1090 was screened with flagellin-specific rabbit antiserum for molecular cloning of the flagellin gene of H. pylori. A positive clone, λHPF4, was obtained and the recombinant antigen expressed from λHPF4 was a fusion protein with the molecular weight of 168kd. Sequence analysis of antigen-encoding DNA showed that an open reading frame composed of 1,536 nucleotides encodes a polypepride with a oredicted molecular size of 54kd. This open reading frame did not show the homology with flaA gene encoding 56kd protein of H. pylori and was confirmed as a unique sequence through homoligy searching. Therefore, the cloned antigen is supposed to be the carboxy-terminal region of the other flagellin protein of H. pylori, flaB, with the molecular weight of 58kd.
Cloning and DNA Sequencing of Flagellin Gene of Helicobacter pylori
RHEE, KWANG-HO,LEE, WON-KON,CHO, MYUNG-JE,DOH, YOUNG-MI,BAIK, SEUNG-CHUL 경상대학교 유전공학연구소 1992 遺傳工學硏究所報 Vol.11 No.-
A λgtll expression library of Helicobacter pylori DNA was screened with flagllin-specific rabbit antiserum for molecular cloning of the flagellin gene of Helicobacter pylori. A positive clone,λHPF4, was obtained and the recombinant antigen expressed from λHPF4 was a fusion protein with the molecular weight of 168kd. Sequence analysis of antigen-encoding DNA ahowed that an open reading frame of 1536 nucletides encodes a polypeptide with a predicted molecular size of 54kd. This open reading frame did not show any homology with flaA gene encoding 56kd protein of Helicobacter pylori amd was confirmed as a uniques sequence through homology searching. The cloned antigen is tentatively supposed to be the carboxy-termonal region of the other flagellin protein of Helicobacter pylori, flaB, with the molecular weight of 58kd.
1987년 한국에서 발생한 렙토스피라병의 혈청역학적 조사
이증훈,박영수,이우곤,김석용,정선식,우준희,박성광,박경희,송영욱,김선영,기정일,최두혁,강성귀,김주완,최강원,김우열,최명식,최인학,장우현,윤성열 대한감염학회 1988 감염 Vol.20 No.3
Human leptospirosis was an unfamiliar disease in Korea until 1984 that outbreak of leptospirosis occurred among farmers and soldiers after field works for harvesting rice. During that time, Lee and Jo confirmed the first Korean cases of leptospirosis by serological test, isolation of causative agent and autopy findings. Afterward several outbreaks occurred also during autumn especially after flood in every years and some characterisitcs of leptospirosis in Korea such as clinical manifestations, serotypes and seroepidemiological features has been revealed by many investigators. Because of the major mode of transmission between rodents and human is by direct contact with leptospiral urine of rodents or contaminated soil by the urine, leptospirosis in Korea has been primarily a disease of person in occupations heavily exposed to contaminated soil or infected urine such as farmer, army and etc. Therefore it seems that leptospirosis is one of the main communicable diseases to be controlled urgently in Korea, for an agricultural people account for almost half of total Korean people. For clarifying the seroepidemiological patterns of human leptospirosis in Korea by sex, month region and main reacting serovars of L. interrogans among acute febrile disease occurred in 1987, 1,773 patient's sers with acute febrile episodes were tested by microagglutination test using 19 representative strains of leptospiral serogroup as antigen. All of those sera were collected from 10 collaborative clinics located in Kyunggi, Kangwon, Chungbuk, Chungnam, Chonbuk, Chonnam province and Seoul. The results wee summerized as follows. 1) Among 1,773 sera of patients with acute febrile episodes, 219 (12.4%) were seropositive to L. interrogans, 487(27.5%) to R. tsutsugamushi, 241(13.6%) to R.typhi and 160(90.0%) to Hantaan virus. 2) Among seropositives to L.interrogans, the male outnumbered the female, 65% and 35%. 3) For age distribution, 26.9% of seropositives to L.interrogans were fifties, 19.6% were forties, 9.1% were sixties, 5.9% were thirties and 4.1% were twenties. 4) Eighty three percent of seropositives had occurred between September and October in 1987 with a peak in September. 5) Main leptospiral serovars reactive to patient's sera were Icterohaemorrhagiae(54.3%), Canicola(31.0%), CH-48(13.2%), Tarassovi(0.9%)and Cynopteri(0.5%). 6) For regional distribution, 65.8% of seropositives to L.interrogans were residents from Chonbuk, 12.3% were Chonnam, 7.3% were Chungnam, 5.5% were Kyunggi and 1.4% were Kangwon.