• 제목/요약/키워드: natural recombination

검색결과 81건 처리시간 0.026초

Mechanisms Underlying the Effects of LPS and Activation-induced Cytidine Deaminase on IgA Isotype Expression

  • Park, Seok-Rae;Kim, Hyun-A;Chun, Sung-Ki;Park, Jae-Bong;Kim, Pyeung-Hyeun
    • Molecules and Cells
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    • 제19권3호
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    • pp.445-451
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    • 2005
  • Activation-induced cytidine deaminase (AID) is needed for Ig class switch recombination (CSR). We explored the effect of LPS on the expression of AID during B cell differentiation, and the role of AID in IgA isotype expression. In normal spleen B cells, LPS increased AID transcription up to 48 h post-stimulation, i.e. around the time of Ig CSR. TGF-${\beta}1$ and AID were required for IgA expression, and LPS contributed to $TGF{\beta}1$-induced IgA production largely by inducing AID. Interestingly, LPS repressed AID transcription in $sIgA^+$ B cells but still stimulated IgA production mainly by increasing the rate of IgA secretion. Our data indicate that LPS contributes to $TGF{\beta}1$-induced IgA isotype expression in at least two ways: by stimulating AID transcription before CSR and by enhancing the IgA secretion rate after CSR.

Enhancement of Photocurrent Generation by C60-encapsulated Single-walled Carbon Nanotubes in Ru-sensitized Photoelectrochemical Cell

  • Lee, Jung-Woo;Park, Tae-Hee;Lee, Jong-Taek;Jang, Mi-Ra;Lee, Seung-Jin;Kim, Hee-Su;Han, Sung-Hwan;Yi, Whi-Kun
    • Bulletin of the Korean Chemical Society
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    • 제33권8호
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    • pp.2689-2693
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    • 2012
  • Single-walled carbon nanotubes (SWNTs) and $C_{60}$-encapsulated SWNTs ($C_{60}@SWNTs$) are introduced to Ru-sensitized photoelectrochemical cells (PECs), and photocurrents are compared between two cells, i.e., an $RuL_2(NCS)_2$/DAPV/SWNTs/ITO cell and an $RuL_2(NCS)_2$/DAPV/$C_{60}@SWNTs$/ITO cell. [L = 2,2'-bipyridine-4,4'-dicarboxylic acid, DAPV = di-(3-aminopropyl)-viologen, and ITO = indium-tin oxide] The photocurrents are increased by 70.6% in the presence of $C_{60}@SWNTs$. To explain the photocurrent increase, the reverse-field emission method is used, i.e., $RuL_2(NCS)_2$/DAPV/SWNTs/ITO cell (or $RuL_2(NCS)_2$/DAPV/$C_{60}@SWNTs$/ITO cell) as an anode and a counter electrode Pt as a cathode in the external electric field. The improved field emission properties, i.e., ${\beta}$ (field enhancement factor) and emission currents in the reverse-field emission with $C_{60}@SWNTs$ indicate the enhancement of the PEC electric field, which implies the improvement of the electron transfer rate along with the reduced charge recombination in the cell.

UVSC of Aspergillus nidulans is a Functional Homolog of RAD51 in Yeast

  • Yoon, Jin-Ho;Seong, Kye-Yong;Chae, Suhn-Kee;Kang, Hyen-Sam
    • BMB Reports
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    • 제34권5호
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    • pp.428-433
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    • 2001
  • A defect in uvsC of Aspergillus nidulans caused high methyl methansulfonate (MMS)-sensitivity, hyporecombination, and a lack of UV induced mutation. The uvsC gene of Aspergillus nidulans shares a sequence similarity with the RAD51 gene of Saccharomyces cerevisiae. In this study, in vitro and in vivo tests were conducted in order to determine whether or not the UVSC protein had functional similarities to RAD51, the recombination enzyme in yeast. The purified recombinant UVSC protein, following expression in Escherichia coli, showed binding activity to single-stranded DNA (ssDNA), when both ATP and magnesium are present. In addition, ATPase activity was also demonstrated and its activity was stimulated in the presence of ssDNA. The UVSC protein that was expressed under the ADH promoter in S. cerevisiae suppressed in part the sensitivity to MMS of the rad51 null mutant. Similarly, when the uvsC cDNA was expressed from the nmt promoter, the MMS sensitivity of the rhp51 null mutant of Schizosaccharomyces pombe was partially complemented. These results indicate that the A. nidulans UVSC protein is a functional homologue of the RAD51 protein.

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Protective Role of Thioredoxin Peroxidase Against Ionizing Radiation

  • Lee, Su-Min;Kim, Sun-Yee;Park, Jeen-Woo
    • BMB Reports
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    • 제31권6호
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    • pp.572-577
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    • 1998
  • A soluble protein from Saccharomyces cerevisiae provides protection against a thiol-containing oxidation system but not against an oxidation system without thiol. This 25-kDa protein acts as a peroxidase but requires the NADPH-dependent thioredoxin system or a thiol-containing intermediate, and was thus named thioredoxin peroxidase. The protective role of thioredoxin peroxidase against ionizing radiation, which generates reactive oxygen species harmful tocellular function, was investigated in wild-type and mutant yeast strains in which the tsa gene encoding thioredoxin peroxidase was disrupted by homologous recombination. Upon exposure to ionizing radiation, there was a distinct difference between these two strains in regard to viability and the level of protein carbonyl content, which is the indicative marker of oxidative damage to protein. Activities of other antioxidant enzymes, such as catalase, superoxide dismutase, glucose-6-phosphate dehydrogenase, and glutathione reductase were increased at 200-600 Gy of irradiation in wild-type cells. However, the activities of antioxidant enzymes were not significantly changed by ionizing radiation in thioredoxin peroxidase-deficient mutant cells. These results suggest that thioredoxin peroxidase acts as an antioxidant enzyme in cellular defense against ionizing radiation through the removal of reactive oxygen species as well as in the protection of antioxidant enzymes.

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마우스 내장 림프조직에서 우세하게 발현되는 IgA Isotype Switching 관련 전사체의 분석 (Preferential Expression of IgA Isotype Switching-associated Transcripts in Mouse Intestinal Lymphoid Tissues)

  • 채병철;전성기;서구영;김현아;김평현
    • IMMUNE NETWORK
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    • 제5권4호
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    • pp.215-220
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    • 2005
  • Background: Transforming growth factor-${\beta}$ (TGF-${\beta}1$) directs class switch recombination (CSR) to IgA isotype, which is a predominant antibody in mucosal surfaces. Although IgA is preferentially committed in mucosal lymphoid tissues, it is not definitely established whether hallmarks of IgA CSR such as IgA germ-line transcripts (GLT ${\alpha}$), post-switch transcripts (PST ${\alpha}$) and circle transcripts (CT ${\alpha}$) are readily expressed in such tissues. Therefore, we compared the expression of these transcripts among mouse Peyer's patches (PP), mesenteric lymph nodes (MLN), and spleen. Methods: Levels of GLTs, PSTs and CTs were measured by RT-PCR in isolated PPs, MLNs and spleen cells. Results: GLT ${\alpha}$ and PST ${\alpha}$ were well expressed in PP and MLN cells but in spleen cells. Similar patterns were observed in the expression of GL ${\gamma}$2b and PST ${\gamma}$2b. On the other hand, these transcripts were only inducible in spleen cells upon stimulated with LPS and TGF-${\beta}1$. In addition, CT${\alpha}$ and CT${\gamma}$2b were detected in PP cells. Conclusion: PP B cells readily express IgA GLT, PST, and CT. Overall expression patterns of these transcripts were similar in MLN cells. Thus, these results suggest that microenvironment of PP and MLN influences spontaneous IgA CSR, which lacks in systemic lymphoid tissues such as spleen.

Newly Identified TLR9 Stimulant, M6-395 Is a Potent Polyclonal Activator for Murine B Cells

  • Park, Mi-Hee;Jung, Yu-Jin;Kim, Pyeung-Hyeun
    • IMMUNE NETWORK
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    • 제12권1호
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    • pp.27-32
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    • 2012
  • Background: Toll-like receptors (TLRs) have been extensively studied in recent years. However, functions of these molecules in murine B cell biology are largely unknown. A TLR4 stimulant, LPS is well known as a powerful polyclonal activator for murine B cells. Methods: In this study, we explored the effect of a murine TLR9 stimulant, M6-395 (a synthetic CpG ODNs) on B cell proliferation and Ig production. Results: First, M6-395 was much more potent than LPS in augmenting B cell proliferation. As for Ig expression, M6-395 facilitated the expression of both TGF-${\beta}1$-induced germ line transcript ${\alpha}$ ($GLT{\alpha}$) and IL-4-induced $GLT{\gamma}1$ as levels as those by LPS and Pam3CSK4 (TLR1/2 agonist) : a certain Ig GLT expression is regarded as an indicative of the corresponding isotype switching recombination. However, IgA and IgG1 secretion patterns were quite different--these Ig isotype secretions by M6-395 were much less than those by LPS and Pam3CSK4. Moreover, the increase of IgA and IgG1 production by LPS and Pam3CSK4 was virtually abrogated by M6-395. The same was true for the secretion of IgG3. We found that this unexpected phenomena provoked by M6-395 is attributed, at least in part, to its excessive mitogenic nature. Conclusion: Taken together, these results suggest that M6-395 can act as a murine polyclonal activator but its strong mitogenic activity is unfavorable to Ig isotype switching.

Occurrence and Evolutionary Analysis of Coat Protein Gene Sequences of Iranian Isolates of Sugarcane mosaic virus

  • Moradi, Zohreh;Nazifi, Ehsan;Mehrvar, Mohsen
    • The Plant Pathology Journal
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    • 제33권3호
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    • pp.296-306
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    • 2017
  • Sugarcane mosaic virus (SCMV) is one of the most damaging viruses infecting sugarcane, maize and some other graminaceous species around the world. To investigate the genetic diversity of SCMV in Iran, the coat protein (CP) gene sequences of 23 SCMV isolates from different hosts were determined. The nucleotide sequence identity among Iranian isolates was more than 96%. They shared nucleotide identities of 75.5-99.9% with those of other SCMV isolates available in GenBank, the highest with the Egyptian isolate EGY7-1 (97.5-99.9%). The results of phylogenetic analysis suggested five divergent evolutionary lineages that did not completely reflect the geographical origin or host plant of the isolates. Population genetic analysis revealed greater between-group than within-group evolutionary divergence values, further supporting the results of the phylogenetic analysis. Our results indicated that natural selection might have contributed to the evolution of isolates belonging to the five identified SCMV groups, with infrequent genetic exchanges occurring between them. Phylogenetic analyses and the estimation of genetic distance indicated that Iranian isolates have low genetic diversity. No recombination was found in the CP cistron of Iranian isolates and the CP gene was under negative selection. These findings provide a comprehensive analysis of the population structure and driving forces for the evolution of SCMV with implications for global exchange of sugarcane germplasm. Gene flow, selection and somehow homologous recombination were found to be the important evolutionary factors shaping the genetic structure of SCMV populations.

수질환경에서 일어나는 항생물질 내성유전자의 전이와 재조합 (Transfer and genetic recombination of antibiotic resistance genes occurring in water environment)

  • 김치경;이성기;김영창
    • 한국미생물·생명공학회지
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    • 제14권3호
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    • pp.245-250
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    • 1986
  • 청주시 무심천의 하천수에서 항생물질에 내성을 나타내는 Gram 음성 세균을 분리하여 수질환경에서 일어나는 R 플라스미드의 전이를 연구하였다. 분리된 균주사이에서 접합에 의한 R 플라스미드의 전이는 실험실 환경에서 1.1$\times$$10^{-6}$-1.2$\times$$10^{-7}$, 하천의 수질환경에서 1.2$\times$$10^{-7}$-1.0$\times$$10^{-9}$으로 나타나, 자연의 수질환경에서도 R 플라스미드의 전이가 일어남을 확인하였다. 또 T-44 균주의 Ap$^{r}$Cm$^{r}$Tc$^{r}$ 플라스미드는 형질전환에 의하여 E. coli HB 101에 1.7$\times$$10^{-6}$의 비율로 전이되었다. 분자의 크기가 약 9.01kb로 측정된 Ap$^{r}$Cm$^{r}$Tc$^{r}$플라스미드 DNA를 제한효소로 처리한 결과 이 플라스비드에는 EcoRI과 BamHI의 절단부위가 각각 하나씩 존재하고 P-stI의 절단부위는 3개가 있었다.

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Genetic Diversity of a Natural Population of Apple stem pitting virus Isolated from Apple in Korea

  • Yoon, Ju Yeon;Joa, Jae Ho;Choi, Kyung San;Do, Ki Seck;Lim, Han Cheol;Chung, Bong Nam
    • The Plant Pathology Journal
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    • 제30권2호
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    • pp.195-199
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    • 2014
  • Apple stem pitting virus (ASPV), of the Foveavirus genus in the family Betaflexiviridae, is one of the most common viruses of apple and pear trees. To examine variability of the coat protein (CP) gene from ASPV, eight isolates originating from 251 apple trees, which were collected from 22 apple orchards located in intensive apple growing areas of the North Gyeongsang and North Jeolla Provinces in Korea, were sequenced and compared. The nucleotide sequence identity of the CP gene of eight ASPV isolates ranged from 77.0 to 97.0%, while the amino acid sequence identity ranged from 87.7 to 98.5%. The N-terminal region of the viral CP gene was highly variable, whereas the C-terminal region was conserved. Genetic algorithm recombination detection (GARD) and single breakpoint recombination (SBP) analyses identified base substitutions between eight ASPV isolates at positions 54 and 57 and position 771, respectively. GABranch analysis was used to determine whether the eight isolates evolved due to positive selection. All values in the GABranch analysis showed a ratio of substitution rates at non-synonymous and synonymous sites (dNS/dS) below 1, suggestive of strong negative selection forces during ASPV CP history. Although negative selection dominated CP evolution in the eight ASPV isolates, SLAC and FEL tests identified four possible positive selection sites at codons 10, 22, 102, and 158. This is the first study of the ASPV genome in Korea.

고등식물의 엽록체 형질전환: 원핵생물과 진맥생물의 조우 (Chloroplast Genetic Transformation in Higher Plants: An Encounter Between Prokaryote and Eukaryote)

  • 정화지;서영배;정원중;민성란;유장렬
    • Journal of Plant Biotechnology
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    • 제33권3호
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    • pp.185-194
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    • 2006
  • 엽록체는 숙주세포에 잡아먹힌 (식균작용) 남세균이 숙주세포와 공생관계를 형성하여 온 것으로 간주된다. 엽록체 게놈은 정적이라고 이해하고 있지만 형질전환을 통하여 상동염기가 도입되면 이와는 반대로 intramolecular homologous recombination에 의해 subgenomic circle을 만드는 등 매우 다이나믹하다는 것이 최근에 증명되고 있다. 고등식물의 엽록체 형질전환은 핵 형질전환에서 기대할 수 없는 여러 이점을 제공한다. 예컨대, transgene의 발현율을 높일 수 있고, transgene들을 polycistronic하게 발현할 수 있으며, 도입된 transgene이 모계유전을 하게 된다는 것 등이다. 담배는 엽록체 형질전환의 모델 식물로 사용되어 왔으나 최근에는 벼, 대두, 면화 등 다른 주요 작물의 형질전환도 가능하게 되었다. 엽록체 형질전환된 작물은 미생물을 이용하여 고부가가치 단백질을 생산하는 생물반응기를 향후 대체할 수 있게 될 것이다.