• 한국미생물·생명공학회지
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• 제27권4호
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• pp.278-285
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• 1999

In order to use heat shock promoter for the detection of toxic substances, dnaK promoter was amplified from E. coli genomic DNA by using a polymerase chain reaction(PCR) followed by sequencing and sub-cloning into the multi-cloning site of the plasmid, pUCD615. The pUCD615 is a broad-host-range vector containing promoterless lux operon originated from V.fischeri. The recombinant plasmid was transfered to E. coli DH5$\alpha$ through electroporation. The recombinant E. coli showed several patterns of bioluminescent responses to ethanol stress. The bioluminescent E. coli also showed responses to other toxic substances including FeK3(CN)6, CdCl2, p-nitrophenol and HgCl2. The increases of RLU(Relative Light Unit) were observed at 100ppm of FeK3(CN)6, 10ppm and 100ppm and 100ppm of CdCl2, 1ppm of 10ppm of p-nitrophenol and at 1ppm of HgCl2.

• 생명과학회지
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• 제13권5호
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• pp.746-750
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• 2003

Lactose operon contains two CRP binding sites at promoter(CRP1 site) and operator(CRP2 site) regions at lac operon. CRP protein can bind to both sites with the different binding affinity. CRP1 site, major CRP binding site, acts the transcription activation with the fully unknown mechanism by binding of CRP. In this study, the binding affinities of CRP1 site and CRP2 site were measured with the fluorescein-labeled oligomers, which contain CRP1 site and the three different spacing sequences between GTGA and TCAC at CRP2 site. Results showed that CRP:cAMP complex bound to CRP1 site 3 times more strongly than CRP2 site and the base spacing between GTGA and TCAC was not the only factor to affect the binding affinity of CRP to CRP2 site.

• Journal of Microbiology
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• 제45권3호
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• pp.234-240
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• 2007

The DcuS-R two-component system of Escherichia coli senses $C_{4}-dicarboxylates$ of the medium and regulates expression of the genes related to utilization of them. It is known that phospho-DcuR induces expression of genes such as the dcuB-fumB operon, the frdABCD operon, and the dctA gene. We analyzed promoters of the dcuS-R operon to elucidate the transcriptional regulation system. We found a novel internal promoter within the dcuS gene that is regulated by the transcriptional regulator, CRP-cAMP, in both aerobic and anaerobic conditions.

• 미생물학회지
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• 제29권4호
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• pp.215-220
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• 1991

MudJ(Km.lac) operon fusions were used in the identification of osmotic-inducible genetic(osi) loci in Salmonella typhimurium. Expression of osi::lacZ(osi5001, 5027) genes were dramatically induced 39-189 fold when the osmolarity was increased. Seven osm::lacZ genes were constituvely expressed under both low and high osmotic strength. The osi5001::lacZ fusion strains showed the enhanced osmotolerance and the reduced expression of the osi5001::lacZ in the presence of 1mM proline or betaine as osmoprotectants. Four osmotic inducible genetic loci were mapped into 36 (YK531), 44 (YK504), 57 (YK501) and 84 (YK528) map unit by testing the cotransduction frequency.

• Journal of Yeungnam Medical Science
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• 제9권2호
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• pp.248-255
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• 1992

S. typhimurium의 세포내생존을 가능케하는 유전자가 다른 세균에서도 존재하는지를 조사하기 위해 8주의 세균주를 사용하였다. phoP-PhoQ operon은 세포내 환경의 자극을 인지하고 그 환경의 적응에 관여하는 유전자의 유전자표현을 조절한다고 알려져 있다. S. typhimurium의 phoP region의 514 basepairs EcoRV DNA fragment를 probe로 dot blot hybridization을 실시하였다. K. pneumoniae, P. aeruginosa, S. marscescens, E. cloacae, S. typhimurium의 phoP/phoQ gene과 유사한 DNA sequence를 가지지 않았으며 E. coli, S. dysenteriae, E. cloacae에서는 세포내 생존가능세균이 아님에도 불구하고 positive signal을 나타내었다. 이상의 결과에서 S. typhimurium외의 세포내 생존가능세균에는 phoP/PhoQ operon이 없다는 것을 알게 되었고 세포내 생존이 가능하지는 않지만 S. typhimurium과 계통발생학적으로 가까운 세균주에서 phoP/phoQ operon이 발견되었다. L. monocytogenes의 세포내 생존에는 phoP/phoQ에 의존하지 않는 어떤 다론 기전이 존재할 것으로 사료된다.

• 한국환경독성학회:학술대회논문집
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• 한국환경독성학회 2001년도 춘계심포지움 및 학술발표회
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• pp.141-141
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• 2001

The arsenic resistance operon from pMH12 in Klebsiella oxytoca contains two regulatory genes. The first open reading frame for arsR extend up to 348 bp and has a translational product corresponding to a protein of 116 amino acid residue polypeptide with a molecular mass of 13 kDa. And the second ORF for arsD extend up to 360 bp and express a protein of 120 amino anid residue polypeptide with 13kDa. (omitted)

• 미생물학회지
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• 제27권3호
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• pp.201-209
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• 1989

Mu dJ(km lac) peron 융합을 이용하여 Salmonella typhimurium에서 산소에 의해 조절되는 여러개의 새로운 유전자좌를 확인하였다. 산소가 있는 조건에서 유도된 9개의 유전자와 무산소조건에서 유도된 13개의 유전자가 분리되었다. 유전자 융합체들은 조절윤전자 oxrA의 control에 근거하여 2classe로 구분되었는데, class I 유전자들은 ozrA에 의해 조절되고, class II 유전자들은이 조절 유전자에 의해 영향을 받지 않았다. 몇몇 anti-lacZ 유전자 융합체들으 최소배지에서 발현되지 않았고, LB상에서나 혹은 CAA 첨가배지에서 그 활성을 나타내었다. 대부분의 유전자 융합체들은 nitrate와 fumarate에서 발현이 억제되었다. ani 유전자 가운데 3개의 유전자는 상호형질도입 빈도에 의해 각각 $59{\pm}0.14$ map unit (YK114), $64{\pm}0.2$ map unit(YK120), and $93{\pm}0.29$ map unit(YK112) 로 결정되었다.

• Journal of Microbiology and Biotechnology
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• 제11권6호
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• pp.1115-1119
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• 2001

Eight Escherichia coli cells with aerobic growth deflects were isolated by the insertion of ${\lambda}placMu53$, a hybrid bacteriophage of ${\lambda}$ and Mu, which created transcriptional fusion to lacZY. Two of these mutants, CLIO and CLl2, were irradiated with UV to obtain specialized transducing phages. The phages that took out the neighboring chromosomal DNA of the related gene responsible for deflective aerobic growth were identified. The in vivo cloned chromosomal sequence revealed that the mutated gene of CLIO was located at min 34.5 on the Escherichia coli linkage map and 1,599,515 on the physical map. The physical map indicated that there were 7 cistrons in the operon. We named this operon oxg10. The promoter sequence of oxg10 exhibited a possible binding site far SoxS, a transcriptional regulator that activates the transcription of various SoxRS regulon genes. Transferring the oxg10:: ${\lambda}placMu53$ mutation into the wild-type strain, RZ4500, resulted in the inhibition of normal aerobic growth, while the salute mutation in strain MO inhibited aerobic cell growth completely. The full operon sequences of oxg10 were cloned from the Excherichia coli genomic library. The mutated gene of CLl2 was identified to be a sucA gene encoding the ${\alpha}$-ketoglutarate dehydrogenase El component in the TCA cycle.

• Journal of Microbiology and Biotechnology
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• 제16권6호
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• pp.988-992
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• 2006

The ptsHI operon from Leuconostoc mesenteroides ssp. mesenteroides SY1 (L. mesenteroides SY1), a strain isolated from kimchi, was cloned and characterized. The ptsH open reading frame (ORF) was 273 bp in size, which can encode a protein of 90 amino acid residues with a molecular weight of 9,212 Da. The pfsI ORF was 1,719 bp in size, which was capable of encoding a protein of 572 amino acids with a molecular mass of 62,549 Da. ptsH and pfsI genes were transcribed as a single transcript of 2.0 kb in size regardless of carbon sources, supporting the operon structure. Although the deduced amino acid sequences of the HPr and EI were highly homologous with those of other Gram-positive bacteria, an additional amino acid (glutamine at the $3^{rd}$ amino acid) was present in HPr from L. mesenteroides SY1. Phosphorylation sites of HPr included the histidine residue ($16^{th}$) and serine residue ($47^{th}$). Mutant HPrs, in which each phosphorylation site was mutated into alanine, were obtained, and phosphorylation with HPr and mutated HPrs showed that HPr was phosphorylated at the serine residue ($47^{th}$) by HPr kinaseiphosphorylase (HPr K/P).

• ALGAE
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• 제17권1호
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• pp.59-68
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• 2002

A phylogenetic affinity between charophytes and embryophytes (land plants) has been explained by a few chloroplast genomic characters including gene and intron (Manhart and Palmer 1990; Baldauf et al. 1990; Lew and Manhart 1993). Here we show that a charophyte, Spirogyra maxima, has the largest operon of angiosperm chloroplast genomes, rpl23 operon (trnⅠ-rpl23-rpl2-rps19-rpl22-rps3-rpl16-rpl14-rps8-infA-rpl36-rps11-rpoA) containing both embryophyte introns, rpl16.i and rpl2.i. The rpl23 gene cluster of Spirogyra contains a distinct eubacterial promoter sequence upstream of rpl23, which is the first gene of the green algal rpl23 gene cluster. This sequence is completely absent in angiosperms but is present in non-flowering plants. The results imply that, in the rpl23 gene cluster, early charophytes had at least two promoters, one upstream of trnⅠ and and another upstream of rpl23, which partially or completely lost its function in land plants. A comparison of gene clusters of prokaryotes, algal chloroplast DNAs and land plant cpDNAs indicated a loss of numerous genes in chlorophyll a+b eukaryotes. A phylogenetic analysis using presence/absence of genes and introns as characters produced trees with a strongly supported clade containing chlorophyll a+b eukaryotes. Spirogyra and embryophytes formed a clade characterized by the loss of rpl5 and rps9 and the gain of trnⅠ (CAU) and introns in rpl2 and rpl16. The analyses support the hypothesis that the rpl23 gene cluster and the rpl2 and rpl16 introns of land plants originated from a common ancestor of Spirogyra and land plants.