• Title/Summary/Keyword: M/BP/1

Search Result 646, Processing Time 0.027 seconds

Characterization of the Nickel Resistance Gene from Legionella pneumophila: Attenuation of Nickel Resistance by ppk (polyphosphate kinase) Disruption in Escherichia coli

  • Hahm, Dae-Hyun;Yeon, Mi-Jung;Ko, Whae-Min;Lee, Eun-Jooh;Lee, Hye-Jung;Shim, In-Sop;Kim, Hong-Yeoul
    • Journal of Microbiology and Biotechnology
    • /
    • v.12 no.1
    • /
    • pp.114-120
    • /
    • 2002
  • A 1,989-bp genomic region encoding nickel resistance genes was isolated from Legionella pneumophila, a pathogen for legionellosis. From a sequencing and computer analysis, the region was found to harbor two structural genes, a nreB-like protein gene (1,149 bp) and a nreA-like protein gene (270 bp), in a row. Both genes exhibited a significant degree of similarity to the corresponding genes from Synechocystis sp. PCC6803 ($54\%$ amino acid sequence identity) and Achromobacter xylosoxidans 31A ($76\%$). The gene was successfully expressed in E. coli MG1655 and conferred a nickel resistance of up to 5 mM in an LB medium and 3 mM in a TMS medium including gluconate as the sole carbon source. E. coli harboring the nickel resistance gene also exhibited a substantial resistance to cobalt, yet no resistance to cadmium or zinc. Since the extracellular concentration of nickel remained constant during the whole period of cultivation, it was confirmed that the nickel resistance was provided by an efflux system like the $Ni^2+$permease (nrsD) of Synechocystis sp. strain PCC6803. Since polyphosphate (poly-P) is known as a global regulator for gene expression as well as a potential virulence factor in E. coli, the nickel resistance of a ppk mutant of E. coli MG 1655 harboring the nickel resistance gene from L. pneumophila was compared with that of its parental strain. The nickel resistance was significantly attenuated by ppk inactivation, which was more pronounced in an LB medium than in a TMS medium.

Production of Xylooligosaccharides by Yeast Cell Surface-Displayed Endoxylanase (효모 세포 표면 발현된 Endoxylanase를 이용한 Xylooligosaccharides의 생산)

  • Kim, Hyun-Jin;Lee, Jae-Hyung;Kim, Yeon-Hee;Nam, Soo-Wan
    • Microbiology and Biotechnology Letters
    • /
    • v.36 no.4
    • /
    • pp.307-313
    • /
    • 2008
  • The yeast surface expression system, pCTXYN (6.8 kb), of Bacillus endoxylanase gene (xynB, 642 bp) was constructed and introduced into Saccharomyces cerevisiae EBY100 cell. The transformed yeast cell showing the highest endoxylanase activity was selected through the active staining of colonies grown on YPDG medium containing xylan. With the yeast transformant, EBY100/pCTXYN, grown on galactose containing medium, it was found that the endoxylanase was successfully displayed on the yeast cell surface and the xylooligosaccharides were efficiently produced from xylan. The most of endoxylanase activity was detected in the cell fraction and reached about 1.9 unit/mL after 48 h cultivation. The optimized conditions for xylooligosaccharides production from xylan were determined as follows: substrate and its concentration, oat spelt xylan 6%; concentration of yeast whole-cell, 5 unit/mL; temperature, $50^{\circ}C$, and reaction time $2{\sim}4\;h$. When the oat spelts xylan and corncob xylan were hydrolyzed by treatment with cell surface-displayed endoxylanase, xylotriose was formed as a main product.

Function of mORF1 Protein as a Terminal Recognition Factor for the Linear Mitochondrial Plasmid pMLP1 from Pleurotus ostreatus

  • Kim, Eun-Kyoung;Roe, Jung-Hye
    • Journal of Microbiology
    • /
    • v.37 no.4
    • /
    • pp.229-233
    • /
    • 1999
  • The mitochondrial plasmid pMLP1 from a white-rot fungus, Pleurotus ostreatus, is a double-stranded DNA containing 381 bp terminal inverted repeat (TIR) whose 5'-ends are covalently bound by terminal proteins. The plasmid contains two major open reading frames (ORFs), encoding putative DNA and RNA polymerases, and a minor ORF encoding a small, highly basic protein. To identify the DNA binding activity that recognizes the TIR region of pMLP1, gel retardation assays were performed with mitochondrial extracts. A specific protein binding to a region between 123 and 248 nt within TIR was observed. We examined whether the gene product of mORF1 bindes to this region specifically. E. coli cell extract which contains an overproduced mORF1 protein formed a complex specific to the region between 123 and 248 nt. Inclusion of mORF1 protein in the specific complex formed between P. ostreatus mitochondrial extract and TIR was confirmed by a supershift assay using polyclonal antibodies against the mORF1 protein. Our result suggest that the product of mORF1 may function as a terminal region recognition factor (TRF), recognizing an internal region in TIR.

  • PDF

Carbonized Rice Excavated at the SORORI Paleolithic Age Site (청원 소로리 구석기유적지 출토 탄화벼)

  • Lee Y.J.;Woo J.Y.;Shu H.S.;Kim J.Y.;Cho Y.K.;Heu M.H.
    • Proceedings of the Korean Society of Crop Science Conference
    • /
    • 1999.05a
    • /
    • pp.120-121
    • /
    • 1999
  • 1. 충북대학교 박물관팀이 충북 청원군 옥산면 소로리에서 구석기시대 유적지를 발굴하는 과정에서 13,010BP로 추정되는 상위토탄층에서 고대형 탄화벼가 출토되었고 36,350BP 이전으로 추정되는 하위토탄층에서 유사벼가 출토되었다. 2. 고대형벼는 11개의 단립형과 1개의 장립형이었다. 장립형벼는 영피표면에 융모가 없고 황금색인(Smootn and gold-hull) 세장립이었다. 그 크기는 지금의 세장한 indica 와 비슷하였다. 3. 유사벼는 영표면에 융모가 있고 깍지(莢)모양으로 되어있는 것(유사벼-1)과 영표면에 융모없이 미끈한것(유사벼-2)으로 2종류가 있었다. 이들의 크기는 모두 벼알과 비슷하였다. 4. DNA 분석결과 고대벼는 지금의 재배종 japonica 나 indica 와는 차이를 보였다. 장립종은 시료가 부족하여 분석하지 못하였다. 유사벼-1은 재배종 벼와 약간 유사한점이 보였으나 유사벼-2는 유사점이 없었다. 5. 고대벼에서는 쌍봉돌기(Bi-peak protuberences)가 있었으나 유사벼-1 과 유사벼-2에서는 이것이 없었다.

  • PDF

Age Dating and Paleoenvironmental Changes of the Kunang Cave Paleolithic Site

  • Yum, Jong-Kwon;Lee, Yung-Jo;Kim, Jong-Chan;Kim, In-Chul;Kim, Ju-Yong
    • The Korean Journal of Quaternary Research
    • /
    • v.17 no.2
    • /
    • pp.145-148
    • /
    • 2003
  • The Kunang cave paleolithic site is located at Tanyang [$N37^{\circ}2'$, $128^{\circ}21'E$], Chungbuk Province, which is in the Central part of the Korean peninsula. The cave is developed at 312 amsl in a karstic mountainous area. The South Han River flows across this region and other caves can also be found near the river. The site was discovered in 1986 and excavated 3 times by the Chungbuk National University Museum until now. The cave was wellpreserved from modem human activities until the first discovery. The full length of the cave is estimated to be ca. 140 m. However, a spacious part up to 11 m from the entrance has been excavated. Eight lithological units are divided over the vertical profile at a depth of 5 m. Each unit is deposited in ascending order as follow: mud layer (Unit 9), lower complex (Unit 8) which is composed of angular blocks and fragments with a muddy matrix, lower travertine layer (Unit 7; flowstone), middle complex (Unit 6; cultural layer) which is composed of fragments with a muddy matrix, middle travertine layer (Unit 5; flowstone), yellowish muddy layer (Unit 4), upper complex (Unit 3; cultural layer) which has a similar composition to Unit 8. the upper travertine layer (Unit 2; flowstone), and finally surface soil layer (Unit 1). The most abundant vestiges in the cultural layers are the animal bones. They are small fractured pieces and mostly less than 3 cm in length. About 3,800 bone pieces from 25 animal species have been collected so far, 90 percent of them belonging to young deers. Previous archaeological study of these bone pieces shows thatprehistoric people occupied the cavenot for permanent dwelling but for temporary shelter during their seasonal hunting activity. More extensive studies of these bones together with pollen analysis are in progress to reconstruct the paleoenvironment of this cave. Only a single date (12,500 BP) obtained from a U-Th measurement of the upper travertine layer was previously available. In spite of the importance of the cave stratigraphy, there was no detail chronological investigation to establish the depositional process of the cultural layers and to understand the periodic structure of the cave strata, alternating travertine floor and complex layers. We have measured five 14C age dating (38900+/-1000, 36400+/-900, 40600+/-1600, more than 51000 and 52000 14C BP) using Seoul National University 14C AMS facility, conducted systematic process of the collagen extraction from bone fragments samples. From the result, we estimate that sedimentation rate of the cave earth is constant, and that the travertine layers, Unit 2 and Unit 3, was formed during MIS 5a(ca. 80 kBP) and MIS 5c (ca. 100 kBP) respectively. The Kunang Cave site is located at Yochonli of the region of Danyang in the mid-eastern part of Korea. This region is compased of limestones in which many caves were found and the Nam-han river flows meanderingly. The excavations were carried out three times in 1986, 1988, and 1998.

  • PDF

The paleo-shoreline and formation age of the 1st marine terrace in Heunghae-eup Pohang City, South Korea : evaluation of the mode and rate of the late Quaternary tectonism (I) (포항시 흥해읍 일대 해안단구 제1면의 구정선 고도와 형성 시기 - 한반도 제4기 후기 지각운동의 양식과 변형률 산출을 위한 연구(I) -)

  • Shin, Jae Ryul;Park, Kyung Geun
    • Journal of the Korean association of regional geographers
    • /
    • v.22 no.3
    • /
    • pp.703-713
    • /
    • 2016
  • This study documents the altitude of paleo-shoreline and formation age of the $1^{st}$ marine terrace emerged around Heunghae-eup Pohang City (South Korea). As a result, the $1^{st}$ terrace representing 10 m of the paleo-shoreline was formed at MIS 5c around 100,000 BP and was influenced repetitive sedimentation of sea-wave till regression of MIS 5a. The result is recognized as a definite truth for the $1^{st}$ terrace in the eastern coast of the Korean peninsula based on synthetic reviews of previous studies and cross-validation of absolute age data. Furthermore, this study deduces a sea stand at MIS 5c from the geomorphological contrast method, but precise determination of paleo-shoreline of the $2^{nd}$ terrace should be required to estimate that of MIS 5c.

  • PDF

Molecular Detection of Mycoplasma felis Infection in a Cat with Respiratory Symptoms

  • Lee, Hyun-A;Hong, Sunhwa;Chung, Yungho;Kim, Okjin
    • Journal of Veterinary Clinics
    • /
    • v.35 no.6
    • /
    • pp.273-275
    • /
    • 2018
  • A 6-month-old male cat was presented for investigation of depression, loss of appetite, dehydration, pale conjunctival mucous membrane, weight loss, fast heart and respiratory rates, nasal discharge and cough. Nasal swabs collected from the studied cat. As the results of bacterial culture with nasal swabs, it was suspected with Mycoplasma spp. Also, Mycoplasma species was detected by the PCR reaction with Mycoplasma genus primers. At species PCR assay, the specimens evaluated for the presence of M. felis, M. arginini, M. gateae, and Acholeplasma laidlawii and the result was visualization of bands from 238 bp in agarose gel 1.5% showing M. felis amplicons in samples. In conclusion, we detected M. felis in a cat with respiratory disease. PCR was able to detect successfully M. felis infection in cats.

Activation and Abnormalities of Cell Cycle Regulating Factor in Head and Neck Squamous Cell Carcinoma Cell Lines: Abnormal Expression of CDKN2 Gene in Laryngeal Squamous Cell Carcinoma (두경부 편평상피세포암 세포주에서 세포주기조절인자의 활성 및 이상 : 후두편평상피세포암에서 종양억제유전자 CDKN2 유전자의 발현이상)

  • Song, Si-Youn;Han, Tae-Hee;Bai, Chang-Hoon;Kim, Yong-Dae;Song, Kei-Won
    • Journal of Yeungnam Medical Science
    • /
    • v.22 no.2
    • /
    • pp.166-182
    • /
    • 2005
  • Background: Cyclin-dependent kinase (CDK) inhibitors are family of molecules that regulate the cell cycle. The CDKN2, a CDK4 inhibitor, also called p16, has been implicated in human tumorigenesis. The CDKN2 inhibits the cyclin/CDK complexes which regulate the transition from G1 to S phase of cell cycle. There is a previous report that homozygous deletion of CDKN2 region on chromosome 9p21 was detected frequently in astrocytoma, glioma and osteosarcoma, less frequently in lung cancer, leukemia and ovarian cancer, but not detected in colon cancer and neuroblastoma. However, little is known about the relationship between CDKN2 and laryngeal cancer. Therefore this study was initiated to investigate the role of CDKN2 in human laryngeal squamous cell carcinoma development.1) Materials and methods: We used 5 human laryngeal carcinoma cell lines whether they have deletions or losses of CDKN2 gene expression by DNA-PCR or RT-PCR, respectively. We examined 8 fresh frozen human laryngeal cancer tissues to detect the loss of heterozygosity (LOH) of CDKN2. PCR was performed by using microsatellite markers of short arm of human chromosome 9 (D9S126, D9S144, D9S156, D9S161, D9S162, D9S166, D9S171, D9S200 and D9SIFNA). For informative cases, allelic loss was scored if the signal of one allele was significantly decreased in tumor DNA when compared to the same allele in normal DNA. Results: The CDKN2 DNA deletion was observed in 3 cell lines. The CDKN2 mRNA expression was observed in only one cell line, which was very weak. LOH was detected in 7 cases (87.5%). Conclusion: These results suggest that CDKN2 plays a role in the carcinogenesis of human laryngeal squamous cell carcinoma.

  • PDF

Isolation and Characterization of Thioredoxin cDNA from Codonopsis lanceolata (S. et Z.) Trautv

  • In, Jun-Gyo;Lee, Bum-Soo;Rho, Yeong-Deok;Yu, Chang-Yeon;Yang, Deok-Chun
    • Korean Journal of Medicinal Crop Science
    • /
    • v.13 no.5
    • /
    • pp.293-297
    • /
    • 2005
  • A thoredoxin (CTRX) gene was cloned and characterized from a full length cDNA library prepared from taproot of three-year old Codonopsis lanceolata. A CTRX was 666 nucleotides long and has an open reading frame of 372 bp with 124 amino acid residues (pI = 4.92). The deduced amino acid sequence of the CTRX matched to the previously reported plant thioredoxin h genes. The deduced amino acid sequence of CTRX exhibited the similarity of 33-67% among previously registered thioredoxin genes. The expression of CTRX in leaves of Codonopsis lanceolata was increased by wounding and 1 mM $H_2O_2$, but decreased by 0.1 mM cadmium.

Human CYP1A2 Promoter Fused-Luciferase Gene Constructs Hardly Respond to Polycyclic Hydrocarbons in Transient Transfection Study in HepG2 Cells

  • Chung, Injae
    • Toxicological Research
    • /
    • v.16 no.2
    • /
    • pp.95-100
    • /
    • 2000
  • In previous study, both constitutive expression and 3-methylcholanthrene (3MC)-mediated elevation of CYP1A2 mRNA were demonstrated in human hepatoma HepG2 cells by reverse transcription-polymerase chain reaction (RT-PCR), suggesting that HepG2 cells would be appropriate for the study of human CYP1A2 regulation(Chung and Bresnick, 1994). Further studies were conducted to determine the basis of this induction phenomenon that is observed in HepG2 cells. Since CYP1A1 gene, another polycyclic hydrocarbon(PH)-inducible gene, is regulated by PHs through their interactions via receptors with cis-elements, the 5'-flanking region of human CYP 1A2 gene was analyzed to search such responsive elements. The promoter activity of various lengths of CYP1A2 gene sequence (-3203/+58bp) was measured in transiently-transfected HepG2 cells by fusion constructs containing the CAT, hGH or luciferase genes as a reporter. This region of the CYP1A2 gene, although containing a XRE, was only weakly responsive (less than 2 fold induction) to 10 nM of TCDD or 1 $\mu$M 3 MC treatment. This small enhancement of promoter activity is inconsistent with the previous observation, i.e., 12 to 14 fold-enhanced CYP1A2 mRNA from 1 $\mu$M 3 MC treated HepG2 cells, suggesting that additional mechanisms would exist for PH-mediated induction of CYP1A2 in these cells.

  • PDF