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Organization and Analysis of the Histidine Biosynthetic Genes from Corynebacterium Glutamicum
Sam Il Jung,Jae Yeon Chun,Sei Heun Yim,Choong Il Cheon,En Sook Song,Soo Suk Lee,Myeong Sok Lee 한국유전학회 2009 Genes & Genomics Vol.31 No.4
Corynebacterium glutamicum, a Gram-positive bacterium, has been widely used for industrial amino acid production. In addition to our previously cloned hisEG and hisHA-impA-hisFI genes, the remaining hisDCB genes were cloned in this study. The entire C. glutamicum histidine biosynthesis genes, when compared with those of other microorganisms, showed high degree of similarities in deduced amino acid sequences but also significant differences in gene organization. Transcription analysis by RT-PCR revealed that C. glutamicum his genes are located and transcribed in two unlinked loci, hisEG and hisDCB-orf1-orf2-hisHA-impA-hisFI. The primer extension analysis showed that the latter his operon starts the transcription at C residue localized 196-bp upstream of the hisD ATG start codon. Genetic analysis in hisD promoter region showed the putative Pribnow boxes, TTTAAT and CAGTAT at 7 and 31 upstream of hisD gene transcription start site. Further analysis revealed Shine-Dalgarno sequence, AGGGAG, at 10-bp upstream of hisD translational start codon. Our result also suggests that the histidine biosynthesis in C. glutamicum is negatively regulated by their end-product, histidine, suggesting the histidine-dependent regulation of his gene transcription.
Molecular Cloning of the Arginine Biosynthetic Genes from Corynebacterium glutamicum
Chun, Jae-Shick,Jung, Sam-Il,Ko, Soon-Young,Park, Mee-Young,Kim, Soo-Young,Lee, Heung-Shick,Cheon, Choong-Ill,Min, Kyung-Hee,Lee, Myeong-Sok The Microbiological Society of Korea 1996 The journal of microbiology Vol.34 No.4
Complementation cloning of the argC, E, B, D, F, and G genes in Corynebacterium glutamicum was done by transforming the genomic DNA library into the corresponding arginine auxotrophs fo Escherichia coli. Recombinant plasmids containing 6.7 kb and 4.8kb fragments complementing the E. coli argB mutant were also able to complement the E. coli argC, E, A, D, and F mutants, indicating the clustered organization of the arginine biosynthetic genes within the cloned DNA fragments. The insert DNA fragments in the recombinant plasmids, named pRB1 AND pRB2, were physically mapped with several restriction enzymes. By further subcloning the entire DNA fragment containing the functions and by complementation analysis, we located the arg genes in the order of ACEBDF on the restriction map. We also determined the DNA nucleotide sequence of the fragment and report here the sequence of the argB gene. When compared to that with the mutant strain, higher enzyme activity of N-acetylglutamate kinase was detected in the extract of the mutant carrying the plasmid containing the putative argB gene, indicating that the plasmid contains a functional argB gene. Deduced amino acid sequence of the argB gene shows 45%, 38%, and 25% identity to that from Bacillus strearothermophilus, Bacillus substilus, and E. coli respectively. Our long term goal is genetically engineering C. glutamicum which produces more arginine than a wild type strain does.
Transcriptional regulation of histidine biosynthesis genes in <i>Corynebacterium glutamicum</i>
Jung, Samil,Chun, Jae-Yeon,Yim, Sei-Heun,Lee, Soo-Suk,Cheon, Choong-Il,Song, Eunsook,Lee, Myeong-Sok Canadian Science Publishing 2010 Canadian journal of microbiology Vol.56 No.2
<P> Corynebacterium glutamicum , a gram-positive bacterium, has been widely used for industrial amino acid production. Corynebacterium glutamicum his genes are located and transcribed in two unlinked loci, hisEG and hisDCB-orf1-orf2-hisHA-impA-hisFI. The latter his operon starts the transcription at the C residue localized 196 bp upstream of the hisD ATG start codon. Our computer-based sequence analysis showed that the region corresponding to the untranslated 5′ end of the transcript, named the hisD leader region, displays the typical features of the T-box transcriptional attenuation mechanism. Therefore, expression of the cat reporter gene under the control of the wild-type or mutated hisD leader regions was tested in multi-copy (pProm and pTer series) and in single-copy (pInt series) systems under conditions of sufficient or limited histidine. Our mutational studies led to the conclusion that the CAU histidine specifier and 5′-UGGA-3′ sequence in the hisD leader region are required for the hisDCB-orf1-orf2-hisHA-impA-hisFI gene regulation. The cat gene expression from the wild-type leader region was negatively regulated by histidine. However, the cat gene expression from mutated leader regions was irresponsive to the level of histidine in the growth medium. Taken together, we propose that a T-box mediated attenuation mechanism is responsible for the gene expression of the hisDCB-orf1-orf2-hisHA-impA-hisFI operon in C. glutamicum. </P>
이종명(Jong Myung Lee),황윤근(Youn Keun Hwang),윤종수(Jong Soo Yun),강천일(Cheon Il Kang),서영익(Young Ik Seo),김능수(Nung Soo Kim),구성모(Seong Mo Koo),조봉기(Bong Kee Cho),강영모(Young Mo Kang),이중기(Choong Ki Lee) 대한내과학회 1997 대한내과학회지 Vol.52 No.1
N/A The number of persons with HIV infection in Korea have increased steadily, total number of HIV infection in Korea were 478 on August, 1995. To investigate the clinicoimmunologic manifestation of AIDS in Korea, we reviewed complete blood counts (CBC), CD4 counts, serum β2-microglobulin level, opportunistic infections and cause of death for 19 AIDS patients who had been admitted or visited at Pusan national university hospital during the period of January, 1990 to August, 1995. 1) The predominant mode of HIV transmission was heterosexual contact(18), other modes of transmission were homosexual contact(1). Clues of diagnosis of HIV infection were routine occupational health examination(14), and opportunistic infection symptoms such as fever, coughing(4). 2) Mean CD4 cell counts(/mm3) were 53±72 totally, 22±27 for 8 dead patients at mean 2 month before, 91±87 for 7 living patients. There were not significant difference(p>0.05). 3) Serum β2-microglobulin(MG;ug/ml) was measured at 12 patients, mean serum β2-MG level was4.8±7.3 totally, 7.1±10.3 for 6 dead patients at mean 1.3 month before, 2.5±0.4 for 6 living patients. There were not significant(p>0.05). 4) At CBC examination, WBC(/mm3) was 5,932±2,899 totally, 5,452±3,436 for 10 dead patients, 6,500 ±2,221 for 9 living patients(p>0.05). Hb(g/dl) was 11,4±2.8 totally, 9.4±1.8 for dead patients, 13.6±1.8 for living patients(p<0.05). Lymphocyte count(/mm) was 1,255±800 totally, 731±424 for dead patients, 1,838716 for living patients(p<0.05). ESR(mm/h) was 72±47 totally, 97±33 for dead patients, 47±47 for living patients(p<0.05). 5) Opportunistic infections had developed at 14 patients, candidiasis 7, pneumocystis carinii pneumonia 5, tuberculosis 3, cytomegalovirus infection 2, herpes zoster 3, toxoplasmosis 1, cryptococcal infection 2, bacterial pneumonia 5, and herpes simplex l. Malignant lymphoma had developed in 1 patient. 6) Mean survival interval from diagnosis of HIV infection to death was 32.8±19.1 months, and the most common cause of death was pneumocystis carinii pneumonia, and other causes of death were meningitis, bacterial pneumonia and AIDS-wasting syndrome. Based on these results, We concluded that CD4 counts, serum β2-microglobulin level, Hb, total lymphocyte count and ESR in AIDS patients are specific laboratory markers of progression and prognosis of AIDS, the most common opportunistic infection was candidiasis, and the most common cause of death in AIDS patients was pneumocystis carinii pneumonia.