• Title/Summary/Keyword: 염색체효소

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Understanding centrosome amplification in cancer: A pathway toward precision-targeted cancer drug development (암의 중심체 증폭 이해를 통한 표적 항암제 개발)

  • Taekyung Kim
    • Journal of Life Science
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    • v.33 no.11
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    • pp.950-955
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    • 2023
  • Cell division is an essential process for the survival and development of living organisms. It is critical that duplicated chromosomes are properly segregated into daughter cells during mitosis. The centrosome is the core organelle that forms the microtubule-organizing center (MTOC), which generates the microtubules that make up the mitotic spindle during cell division. The centrosome is also involved in cell signaling and motility. In normal cells, there is one centrosome in G1 that replicates into two in the S phase and matures through G2. During the M phase, duplicated centrosomes move to both ends of the cell, and spindle microtubules that are generated from MTOC move the chromosome to both ends. The cells then split into two to complete the cell division. However, a phenomenon called centrosome amplification (CA), in which the number of centrosomes is higher than normal, is common in cancer cells and can lead to chromosome instability (CIN). This paper discusses the process of centrosome replication and the role of PLK4 in this process. The possible consequences of centrosome amplification and how the PLK4 inhibitor may be able to treat certain types of cancer cells, such as breast cancer and neuroblastoma, will also be discussed.

Transient neonatal diabetes mellitus with macroglossia diagnosed by methylation specific PCR (MS-PCR) (메틸화 특이 PCR로 진단된 거설증을 동반한 일과성 신생아 당뇨병)

  • Jin, Hye Young;Choi, Jin-Ho;Kim, Gu-Hwan;Yoo, Han-Wook
    • Clinical and Experimental Pediatrics
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    • v.53 no.3
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    • pp.432-436
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    • 2010
  • Transient neonatal diabetes mellitus (TNDM) has been associated with paternal uniparental isodisomy of chromosome 6, paternally inherited duplication of 6q24, or a methylation defect at a CpG island of the ZAC or HYMAI gene. We experienced a case of TNDM in which the patient presented with hyperglycemia, macroglossia, and intrauterine growth retardation, caused by a paternally derived HYMAI. An 18-day-old female infant was admitted to the hospital because of macroglossia and recurrent hyperglycemia. In addition to the macroglossia, she also presented with large fontanelles, micrognathia, and prominent eyes. Serum glucose levels were 200-00 mg/dL and they improved spontaneously 2 days after admission. To identify the presence of a maternal methylated allele, bisulfite-treated genomic DNA from peripheral blood was prepared and digested with BssHII after polymerase chain reaction (PCR) amplification with methylation-specific HYMAI primers. PCR and restriction fragment length polymorphism analysis showed that the patient had only the paternal origin of the HYMA1 gene. TNDM is associated with a methylation defect in chromosome 6, suggesting that an imprinted gene on chromosome 6 is responsible for this phenotype.

Construction of Pretense-defective Mutant of Bacillus subtilis by Homologous DNA Recombination (상동성 유전자재조합을 이용한 단백질분해효소 비생산 바실러스균주의 구축)

  • Lee, Jin-Tae;An, Bong-Jeun
    • Food Science and Preservation
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    • v.7 no.4
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    • pp.414-417
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    • 2000
  • Competent cell transformation of B. subtilis AC819 was carried out using phenotypic protease-defective(Npr-) DNA of B. subtilis MT-2. An obtained transformant, designated B. subtilis HL-1, was obtained by homologous DNA recombination. Phenotypes of B. subtilis HL-1 were characterized histidine requirement streptomycin-resistance, tetracyclin resistance and non-producing protease. Protoplast transformation frequency of B. subtilis HL-1 by plasmid pUB110 was higher than that of B. subtilis MT-2. From this result, B. subtilis HL-1 is useful for protease gene transformation and thermostable protease gene cloning as a host.

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Expression of Glucose Isomerase Gene from Bacillus licheniformis in Escherichia coli. (Bacillus licheniformis 포도당 이성화 효소 유전자의 Excherichia coli에 발현)

  • 신명교;고영희
    • Korean Journal of Microbiology
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    • v.23 no.2
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    • pp.138-146
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    • 1985
  • A Bacillus licheniformis ATCC31667 gene coding for a glucose isomerase has been cloned and expressed in glucose isomerase negative mutant of Escherichia coli. A recombinant plasmid, constructed by ligation of a EcoRI fragment of B.licheniformis chromosomal DNA to vector plasmid pBR322, was expressed glucose isomerase positive in E.coli LE392-6 with growth on minimal medium containing xylose as a sole carbon source. This recombinant plasmid, designated pBGI6, had the insery of 4.1Kb of Bacillus gene in EcoRI site, and restriction map of the plasmid was established. The plasmid pBG16 was very stable after 10days of serial transfer to a fresh medium. The activity of glucose isomerase from the transformed cell containing pBGI6 was increased about 20 fold than its wild type of host.

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MAPK Activity in Porcine Oocytes Maturing InVitro (유사분열 활성화 단백질 효소가 돼지난자의 체외성숙에 미치는 영향)

  • Lee, Jae-Dal
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.11 no.6
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    • pp.2124-2128
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    • 2010
  • In this study, we determined effects of the mitogen-activated protein kinase (MAPK) inhibitor, U0126 on meiotic maturation, microtubule organization and actin filament assembly in the porcine oocyte. The phosphorylated MAPK was first detected at 12 h after the initiation of maturation cultures, fully activated at 24h, and remained until metaphase II. Treatment of germinal vesicle (GV) stage oocytes with $20{\mu}M$ U0126 completely blocked MAPK phosphorylation, but germinal vesicle breakdown (GVBD) was normally proceeded. However, the oocytes didn‘t progress to the metaphase I. The inhibition of MAPK resulted in abnormal spindles. In oocytes treated with U0126 after GVBD, polar body extrusion was normal, but the organization of the metaphase plate and chromosome segregation were abnormal. In conclusion, MAPK activity plays an important regulatory role in GV chromatin configuration and meiotic progress in porcine oocyte maturation.

Construction of an Expression Vector System with the GAP Promoter in Saccharomyces cerevisiae (효모, Saccharomyces cervisiae의 GAP 유전자를 이용한 발현 벡터계의 개발)

  • 황요일;서애란;심상국;정동효
    • Microbiology and Biotechnology Letters
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    • v.19 no.6
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    • pp.568-574
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    • 1991
  • The cloned glyceraldehyde-3-phosphate dehydrogenase (GAP) gene of Saccharomyces cereviszae (Holland et al., 1983) has been characterized. Based on the communication, we have also cloned 2.1 kb CAP DNA fragment and modified this fragment as a portable promoter. Two yeast expression vectors, one is YCp type vector being maintained at low copy number (1 or 2) and the other is YEp type vector at high copy number, have been constructed with the GAP promoter and the PH05' gene as a reporter. Our plasrnids were introduc,ed into S. cerevisiae HY-1, which has been improved. The $Trp^+$ transformants expressed APase activity efficiently and showed high level of PH05' transcripts.

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MLPA Applications in Genetic Testing (유전자진단에 있어서 Multiplex Ligation Dependent Probe Amplification (MLPA)의 이론과 실제)

  • Kim, Gu-Hwan;Lee, Beom-Hee;Yoo, Han-Wook
    • Journal of Genetic Medicine
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    • v.6 no.2
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    • pp.146-154
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    • 2009
  • Multiplex ligation dependent probe amplification (MLPA) is a PCR-based method to detect gene dosage. Since its introduction, MLPA has been used to test a large number of genes for major deletions or duplications. Genetic testing, as a diagnostic tool for genetic disease, has been used primarily to identify point mutations, including base substitutions and small insertions/deletions, using PCR and sequence analysis. However, it is difficult to identify large deletions or duplications using routine PCR- gel based assays, especially in heterozygotes. The MLPA is a more feasible method for identification of gene dosage than another routine PCR-based methods, and better able to detect deleterious deletions or duplications. In addition to detection of gene dosage, MLPA can be applied to identify methylation patterns of target genes, aneuploidy during prenatal diagnoses, and large deletions or duplications that may be associated with various cancers. The MLPA method offers numerous advantages, as it requires only a small amount of template DNA, is applicable to a wide variety of applications, and is high-throughput. On the other hand, this method suffers from disadvantages including the possibility of false positive results affected by template DNA quality, difficulties identifying SNPs located in probe sequences, and analytical complications in quantitative aspects.

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Mapping of UV-B sensitive gene in Arabidopsis by CAPS markers (CAPS marker에 의한 Arabidopsis의 자외선 B 감수성 유전자 지도작성)

  • 박홍덕;김종봉
    • Journal of Life Science
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    • v.12 no.6
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    • pp.715-720
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    • 2002
  • A mutant Arabidopsis thaliana which is very sensitive to Ultraviolet-B(UV-B) radiation has been isolated by ethylmethane sulfonate(EMS) mutagenesis. Genetic cross proved the UV-sensitive gene(uvs) to segregate as a single Mendelian locus. For mapping of uvs, we crossed Arabidopsis thaliana Lansberg with uvs plant(Columbia), and made F2 plants by F1 selfcross. We designed 10 kinds of CAPS marker primers. Each primers amplifies a single mapped DNA sequence from uvs and Lansberg erecta ecotyres. Also identified was at least one restriction endonuclase for each of these PCR product that generates ecotype-specific digestion pattern. We got crossing over value of UB-sensitivity and each CAPS marker which located on different chromosome arm. The value of crossing over showed that uvs was linked to LFY3 which was on chromosome 5.

Molecular Cloning of Serratia rnarcescens Metalloprotease Gene into Escherichia coli (Serratia marcescens Metalloprotease 유전자의 대장균에로의 클로닝)

  • 김기석;이창원;이상열;이병룡;신용철
    • Microbiology and Biotechnology Letters
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    • v.20 no.3
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    • pp.280-288
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    • 1992
  • Molecular cloning of metalloprotease gene from Serratia marcescens ATCC 21074 into Escherichia coli JM109 was carried out. Chromosomal DNA of S. marcescens was completely digested with Hind111 and southern hybridization with a synthetic oligonucleotide probe revealed that a 50 KD metalloprotease gene was contained in 4.0 Kb chromosomal DNA fragment, 4.0 Kb chromosomal DNA fragments eluted from agarose gel were ligated with pUC19 and transformed into E. coli JM109. Nine positive clones were obtained from about $1\times 10^3$ transformants by colony hybridization. Their recombinant plasmids, pSPl and pSP2 have same chromosomal DNA fragments in pUC19 in opposite-orientations. When cloned metalloprotease gene was expressed in E. coli, about 52 KD precursor protein of metalloprotease was detected by western blot analysis from E. coli harboring a recombinant plasmid pSP2. Plasmid pSP2 showed no protease activities in E. coli but overproduced the active metalloprotease in S. rnarcescens ATCC 27117.

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Uptake of Isolated Lyophyllum ulmarium Chromosomes by Ganoderma applanatum Protoplasts (잔나비걸상버섯 원형질체(原形質體)에 의(依)한 만가닥버섯 염색체(染色體)의 섭입(攝入))

  • Yoo, Young-Bok;You, Chang-Hyun;Chang, Kwon-Yawl
    • The Korean Journal of Mycology
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    • v.16 no.4
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    • pp.247-252
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    • 1988
  • The uptake of isolated Lyophyllum ulmarium chromosomes by Ganoderma applanatum protoplasts was induced with polyethylen $glycol+CaCl_2+glycine$. Uptake products of chromosomes by protoplasts showed micro-and macrotransgenome type. The former was slowly growing and unstable, the latter was outgrowing and stable mycelial colonies which made thick hyphae of width and segregation of mycelial colony on GCM containing benomyl. A comparison of macrotransgenome type was using isozyme analysis of esterase. The enzyme pattern of two transformants was distinct in position and quantity compared with parents.

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