• Title/Summary/Keyword: gene rearrangement

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Detection of DNA Rearrangement in Rice Using a Cosmid Library

  • Mun, Eun-Pyo;Nahm, Baek-Hie
    • Animal cells and systems
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    • v.1 no.4
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    • pp.629-635
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    • 1997
  • Previously we reported the migration and rearrangement of a chloroplast gene cluster into mitochondria. The exact genomic locations of the clusters, modes of the gene rearrangement and mechanisms of the interorganellar migration of the clusters have yet to be understood. The detailed analysis needs to include a larger region of DNA surrounding each cluster. To study DNA rearrangement and migration in more detail a cosmid library was constructed using the total rice genomic DNA including nuclear, chloroplast and mitochondrial DNA. From this cosmid library, a sub-library was obtained by selecting the clones hybridized to various regions of chloroplast DNA. According to the hybridization pattern 136 clones from the sub-library were classified into 29 groups. Detailed analysis of these clones revealed that in addition to authentic chloroplast DNA, the clones contain its homologs resulted from rearrangement and mutation. We analyzed two clones in detail, which contain different rp12 homologs resulted from rearrangement and/or migration, respectively.

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Gene Reangement through 151 bp Repeated Sequence in Rice Chloroplast DNA (벼 엽록체 DNA내의 151 bp 반복염기서열에 의한 유전자 재배열)

  • Nahm, Baek-Hie;Kim, Han-Jip
    • Applied Biological Chemistry
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    • v.36 no.3
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    • pp.208-214
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    • 1993
  • To investigate the gene rearrangement via short repeated sequences in chloroplast DNA, the pattern of heterologous gene clusters containing the 151 bp repeated sequence with the development of plastid was compared in rice and the homologous gene clusters from various plant sources were searched for comparative analysis. Southern blot analysis of rice DNA using rp12 gene containing 151 bp repeated sequence as a probe showed the presence of heterologous gene clusters. Such heterologous gene clusters varied with the development of plastid. Also it was observed that the heterologous gene clusters were observed in all of the rice cultivars used in this work. Finally the comparative analysis of DNA sequence of the homologous gene clusters from various plants showed the evolutionary gene rearragngement via short repeated sequence among plants. These results suggest the possible relationship between the plastid development and gene rearrangement through short repeated sequences.

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Heterogeneity of Chloroplast DNA in Rice (벼 엽록체 DNA의 이질성)

  • 남백희;문은표
    • Proceedings of the Botanical Society of Korea Conference
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    • 1987.07a
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    • pp.391-401
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    • 1987
  • Plant chloroplast DNA exists as an unique circular structure in which large single copy(LSC) region and small single copy (SSC) region are separated by large inverted repeat sequences (IRS). It has been known that the unique existence of inverted repeat sequences in chloroplast DNA has no relation with the stability of the chloroplast DNA, but causes the inversion between inverted repeat its biological significance has not been understood so far. In rice, several gene clusters have been cloned and sequenced which contain ribulose-5-biophosphate car-boxylase large subunit (rbcL). Especially, one rbcL gene is linked with rp12 gene which is located in the IRS region in one of the gene clusters. By comparison of nucleotide sequence, the two genes are found to be linked through 151 bp repeat sequence which is homologous to the rp123 gene in IRS region. The repeat sequence is found to be located 3' downstream of rfcL gene and near psbA gene in LSC region. The existence of these repeat sequences and the presence of gene clusters caused by the gene rearrangement thorough the repeat sequence provide a possible which is found to be dispersed chloroplast DNA provide the model system to explaine the heterogeneity of the chloroplast DNA in rice in term of gene rearrangement.

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Evidence for VH Gene Replacement in Human Fetal B Cells

  • Lee, Jisoo;Cho, Young Joo;Lipsky, Peter E.
    • IMMUNE NETWORK
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    • v.2 no.2
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    • pp.79-85
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    • 2002
  • Background: In contrast to evidences of Ig H chain receptor editing in transformed cell lines and transgenic mouse models, there has been no direct evidence that this phenomenon occurs in human developing B cells. Methods: $V_HDJ_H$ rearrangements were obtained from genomic DNA of individual $IgM^-$ B cells from liver and $IgM^+B$ cells from bone marrow of 18 wk of gestation human fetus by PCR amplification and direct sequencing. Results: We found three examples of H chain receptor editing from $IgM^+$ and $IgM^-human$ fetal B cells. Two types of $V_H$ replacements were identified. The first involved $V_H$ hybrid formation, in which part of a $V_H$ gene from the initial VDJ rearrangement is replaced by part of an upstream $V_H$ gene at the site of cryptic RSS. The second involved a gene conversion like replacement of CDR2, in which another $V_H$ gene donated a portion of its CDR2 sequence to the initial VDJ rearrangement. Conclusion: These data provide evidence of receptor editing at the H chain loci in developing human B cells, and also the first evidence of a gene conversion event in human Ig genes.

Gametophytic Abortion in Heterozygotes but Not in Homozygotes: Implied Chromosome Rearrangement during T-DNA Insertion at the ASF1 Locus in Arabidopsis

  • Min, Yunsook;Frost, Jennifer M.;Choi, Yeonhee
    • Molecules and Cells
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    • v.43 no.5
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    • pp.448-458
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    • 2020
  • T-DNA insertional mutations in Arabidopsis genes have conferred huge benefits to the research community, greatly facilitating gene function analyses. However, the insertion process can cause chromosomal rearrangements. Here, we show an example of a likely rearrangement following T-DNA insertion in the Anti-Silencing Function 1B (ASF1B) gene locus on Arabidopsis chromosome 5, so that the phenotype was not relevant to the gene of interest, ASF1B. ASF1 is a histone H3/H4 chaperone involved in chromatin remodeling in the sporophyte and during reproduction. Plants that were homozygous for mutant alleles asf1a or asf1b were developmentally normal. However, following self-fertilization of double heterozygotes (ASF1A/asf1a ASF1B/asf1b, hereafter AaBb), defects were visible in both male and female gametes. Half of the AaBb and aaBb ovules displayed arrested embryo sacs with functional megaspore identity. Similarly, half of the AaBb and aaBb pollen grains showed centromere defects, resulting in pollen abortion at the bi-cellular stage of the male gametophyte. However, inheritance of the mutant allele in a given gamete did not solely determine the abortion phenotype. Introducing functional ASF1B failed to rescue the AaBb- and aaBb-mediated abortion, suggesting that heterozygosity in the ASF1B gene causes gametophytic defects, rather than the loss of ASF1. The presence of reproductive defects in heterozygous mutants but not in homozygotes, and the characteristic all-or-nothing pollen viability within tetrads, were both indicative of commonly-observed T-DNA-mediated translocation activity for this allele. Our observations reinforce the importance of complementation tests in assigning gene function using reverse genetics.

Conjugal transfer and fate of the genetically engineered $Km^{r}$ gene in freshwater environments (유전자 조작기법으로 변형시킨 $Km^{r}$ 유전자의 담수 환경에서의 전이 및 행방)

  • 김치경;이성기
    • Korean Journal of Microbiology
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    • v.28 no.3
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    • pp.219-228
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    • 1990
  • A kanamycin resistance($Km^r$) gene was studied for its transfer in natural freshwater environments by using the natural bacterial isolate(M1) of DK1 and the DKC601 strain, $Km^r$ plasmid of which was genetically engineered from the NI strain. The transfer frequency ofthe $Km^r$ gene and rearrangement of the $Km^r$ plasmid were compared between the gnetically engineered microorganism(GEM) and the NI parental strain by conjugation with the same recipient strain. The transfer frequency of the $Km^r$ gene was about $9.1\times 10^{-12}-1.8\times 10^{-11}$ in both the GEM and NI strains at 5 to $10^{\circ}C$, but the frequency of the NI was about 10 times higher than that of the GEM at 20 to $30^{\circ}C$. The $Km^r$ plasmid in the transconjugants obtained by conjugation of the NI with the MY1 strain as a ricipient showed alot of rearrangement, but the $Km^r$ plasmid transferred from the GEM was stable without alteration of its size. When the MT2 strain was used as a recipient, however, such a rearrangement of the $Km^r$ plamid was observed in the transconjugants obtained from the GEM as well as the NI strain. In those transconjugants obtained from different mating pairs and water environments, the plasmid were appeared to decrease in their number as the period of conjugation time was prolonged, but only the $Km^r$ plasmid transferred from the GEM kept having its size of 52kb. Therefore, the $Km^r$ gene was transferred at the same rate from the GEM and NI strains in natural freshwater environment, but the gene of the GEM strain was more stable than the NIduring conjugation and the $Km^r$ plasmid was rearranged by changing the recipient strain for conjugation in any water environments.

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Detection of the BCR/abl Gene Rearrangement by Reverse Transcriptase Based Polymerase Chain Reaction

  • Lee, Kyung-Ok;Park, Young-Suk;Kim, Yong-Woo;Han, Jung-A;Kim, Yoon-Jung
    • BMB Reports
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    • v.29 no.3
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    • pp.241-247
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    • 1996
  • The Philadelphia (Ph) chromosome is the single most intensively studied chromosome alteration characterizing a human malignancy. The specific genetic alteration of chronic myelogenous leukemia (CML) is the formation of the BCR/abl fusion gene in leukemic cells. The presence of the BCR/abl gene has important diagnostic and prognostic implications in CML. The detection of BCR/abl transcripts by reverse transcriptase based polymerase chain reaction (RT-PCR) was investigated in patients with CML in whom the Ph chromosome abnormality was documented by cytogenetic analysis. In a total of 68 CML patient cases, the Ph chromosome was found in 53 cases (77.9%) by cytogenetic analysis. On the other hand, sixty two cases (91.2%) were detected to have BCR/abl gene rearrangement Of these, b3a2 was 44 cases (64.7%) and b2a2 was 17 cases (25,0%). There was one case with both b3a2 and b2a2 (1.5%). Of the fifteen cases of Ph chromosome negative by cytogenetic anlaysis, the BCR/abl gene was observed in nine cases, The results of BCR/abl fusion gene confirmed by the direct sequencing method correlated well with PCR analysis, The amplified PCR products were detected by $1{\times}10^{-5}$ dilutions. In conclusion, PCR technique is sensitive, rapid and relatively simple for a laboratory test in detecting the BCR/abl fusion gene with CML regardless of the result of cytogenetic analysis.

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Acceleration of X-chromosome gene order evolution in the cattle lineage

  • Park, Woncheoul;Oh, Hee-Seok;Kim, Heebal
    • BMB Reports
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    • v.46 no.6
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    • pp.310-315
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    • 2013
  • The gene order on the X chromosome of eutherians is generally highly conserved, although an increase in the rate of rearrangement has been reported in the rodent lineage. Conservation of the X chromosome is thought to be caused by selection related to maintenance of dosage compensation. However, we herein reveal that the cattle (Btau4.0) lineage has experienced a strong increase in the rate of X-chromosome rearrangement, much stronger than that previously reported for rodents. We also show that this increase is not matched by a similar increase on the autosomes and cannot be explained by assembly errors. Furthermore, we compared the difference in two cattle genome assemblies: Btau4.0 and Btau6.0 (Bos taurus UMD3.1). The results showed a discrepancy between Btau4.0 and Btau6.0 cattle assembly version data, and we believe that Btau6.0 cattle assembly version data are not more reliable than Btau4.0.

Detection of Canine Lymphoma by the Amplification of Antigen Receptor Gene Rearrangements (재배열 항원 수용체 유전자 증폭을 통한 개 림프종의 진단)

  • Yu, Do-Hyeon;Li, Ying-Hua;Lee, Jong-Hyun;Noh, Dong-Ho;Song, Ru-Hui;Lee, Mi-Jin;Choi, Ul-Soo;Park, Jin-Ho
    • Journal of Veterinary Clinics
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    • v.26 no.5
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    • pp.419-422
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    • 2009
  • We performed the PARR (PCR to detect antigen receptor rearrangements) test on DNA isolated from twelve archival canine cytological slides including nine lymphoma, two reactive lymphocytes and one sample from Ehrlichia canis infected dog. As a result, our PCR control gene, $C{\mu}$, was successfully amplified from all of the DNA samples. Six out of nine lymphoma samples showed a clonal rearrangement of immunoglobulin gene whereas three samples did a clonal rearrangement of T cell receptor gamma ($TCR{\gamma}$) gene. However, we observed no visible or clear bands from PCR conducted using our antigen receptor rearrangement primers on DNA from a reactive lymphoid cell proliferation used as a negative control. False-positive amplification in $TCR{\gamma}$ gene was observed only in one sample from E. canis infection. The use of archival cytological specimens demonstrated in this study offers potential advantages for cost-effective specimen acquisition and efficient high-fidelity DNA analysis.