• Title/Summary/Keyword: natural recombination

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Analysis of Natural Recombination in Porcine Endogenous Retrovirus Envelope Genes

  • Lee, Dong-Hee;Lee, Jung-Eun;Park, Nu-Ri;Oh, Yu-Kyung;Kwon, Moo-Sik;Kim, Young-Bong
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.585-590
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    • 2008
  • Human tropic Porcine Endogenous Retroviruses (PERVs) are the major concern in zoonosis for xenotransplantation because PERVs cannot be eliminated by specific pathogen-free breeding. Recently, a PERV A/C recombinant with PERV-C bearing PERV-A gp70 showed a higher infectivity (approximately 500-fold) to human cells than PERV-A. Additionally, the chance of recombination between PERVs and HERVs is frequently stated as another risk of xenografting. Overcoming zoonotic barriers in xenotransplantation is more complicated by recombination. To achieve successful xenotransplantation, studies on the recombination in PERVs are important. Here, we cloned and sequenced proviral PERV env sequences from pig gDNAs to analyze natural recombination. The envelope is the most important element in retroviruses as a pivotal determinant of host tropisms. As a result, a total of 164 PERV envelope genes were cloned from pigs (four conventional pigs and two miniature pigs). Distribution analysis and recombination analysis of PERVs were performed. Among them, five A/B recombinant clones were identified. Based on our analysis, we determined the minimum natural recombination frequency among PERVs to be 3%. Although a functional recombinant envelope clone was not found, our data evidently show that the recombination event among PERVs may occur naturally in pigs with a rather high possibility.

Development of a novel genetic assay for telomere recombination in Saccharomyces cerevisiae (효모에서 텔로미어 재조합을 관찰하기 위한 새로운 유전학적 연구방법의 개발)

  • Kim, Min-Kyu;Bae, Sung-Ho
    • Korean Journal of Microbiology
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    • v.52 no.1
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    • pp.116-119
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    • 2016
  • Stable maintenance of telomere is required for cell proliferation and survival. Although telomerase is the primary means for telomere maintenance, recombination is another important pathway to maintain telomeres. In this study, we developed a genetic assay for telomere recombination using the internal $TG_{1-3}$ repeats present in subtelomeric regions of yeast. The recombination frequencies were dependent on the presence of the internal $TG_{1-3}$ repeats. PCR amplification of the regions near URA3 and CAN1 markers using genomic DNA isolated from $FOA^rCan^r$ colonies indicated that each isolate had lost the chromosome end including the markers. In addition, the recombination frequencies increased with longer internal $TG_{1-3}$ repeats. Our results suggest that the $FOA^rCan^r$ colony formation is the consequence of recombination between the internal and terminal $TG_{1-3}$ repeats.

Methylated-UHRF1 and PARP1 interaction is critical for homologous recombination

  • Hahm, Ja Young;Kang, Joo-Young;Park, Jin Woo;Jung, Hyeonsoo;Seo, Sang-Beom
    • BMB Reports
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    • v.53 no.2
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    • pp.112-117
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    • 2020
  • A recent study suggested that methylation of ubiquitin-like with PHD and RING finger domain 1 (UHRF1) is regulated by SET7 and lysine-specific histone demethylase 1A (LSD1) and is essential for homologous recombination (HR). The study demonstrated that SET7-mediated methylation of UHRF1 promotes polyubiquitination of proliferating cell nuclear antigen (PCNA), inducing HR. However, studies on mediators that interact with and recruit UHRF1 to damaged lesions are needed to elucidate the mechanism of UHRF1 methylation-induced HR. Here, we identified that poly [ADP-ribose] polymerase 1 (PARP1) interacts with damage-induced methylated UHRF1 specifically and mediates UHRF1 to induce HR progression. Furthermore, cooperation of UHRF1-PARP1 is essential for cell viability, suggesting the importance of the interaction of UHRF1-PARP1 for damage tolerance in response to damage. Our data revealed that PARP1 mediates the HR mechanism, which is regulated by UHRF1 methylation. The data also indicated the significant role of PARP1 as a mediator of UHRF1 methylation-correlated HR pathway.

ILLEGITIMATE RECOMBINATION MEDIATED BY MAMMALIAN DNA TOPOISOMERASE II

  • BAE YOUNG-SEUK;IKEDA HIDEO
    • Proceedings of the Microbiological Society of Korea Conference
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    • 1991.04a
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    • pp.82-96
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    • 1991
  • To understand the mechanism of illegitimate recombination in mammalian cells, we have examined the recombination role of DNA topoisomerase II (Topo II ). We found that purified calf thymus Topo II mediates recombination between two phage $\lambda$ DNA molecules in an in vitro system. The enzyme mainly produced a linear monomer recombinant DNA that can be packaged in vitro. Novobiocin and anti-calf thymus Topo II antibody inhibit this ATP-dependent recombination. The recombinant molecules contain duplications or deletion, and most crossovers take place between nonhomologous sequences of $\lambda$ DNA, as judged by the sequences of recombination junctions. In order to study the effects of Topo II on illegitimate recombination in mammalian cells, we have developed a new shuttle vector, pNKl, which contains three bacterial genes, amp(APR), galK and neo($Km^R$). Using this system, we have shown that a Topo II inhibitor, VM26, stimulated deletion formation in pNK1 DNA in monkey COS1 cells. Both in vitro and in vivo results suggest that Topo II participates in illegitimate recombination in mammalian cells.

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Formation of a New Solo-LTR of the Human Endogenous Retrovirus H Family in Human Chromosome 21

  • Huh, Jae-Won;Kim, Dae-Soo;Ha, Hong-Seok;Kim, Tae-Hong;Kim, Wook;Kim, Heui-Soo
    • Molecules and Cells
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    • v.22 no.3
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    • pp.360-363
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    • 2006
  • Human endogenous retroviruses (HERVs) contribute to various kinds of genomic instability via rearrangement and retrotransposition events. In the present study the formation of a new human-specific solo-LTR belonging to the HERV-H family (AP001667; chromosome 21q21) was detected by a comparative analysis of human chromosome 21 and chimpanzee chromosome 22. The solo-LTR was formed as a result of an equal homologous recombination excision event. Several evolutionary processes have occurred at this locus during primate evolution, indicating that mammalian-wide interspersed repeat (MIR) and full-length HERV-H elements integrated into hominoid genomes after the divergence of Old World monkeys and hominoids, and that the solo-LTR element was created by recombination excision of the HERV-H only in the human genome.

Dynamic Response of Charge Recombination from Post-Annealing Process in Organic Solar Cell Using Intensity Modulated Photovoltage Spectroscopy

  • Jeong, Hanbin;Yun, Suk-Jin;Lee, Jae Kwan
    • Journal of Integrative Natural Science
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    • v.9 no.4
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    • pp.275-280
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    • 2016
  • Intensity modulated photovoltage spectroscopy (IMVS) analysis of organic solar cells (OSCs) with a bulk-heterojunction (BHJ) film composed of P3HT and $PC_{61}BM$ was performed. The dynamic response of charge recombination by the post-annealing approach in $P3HT/PC_{61}BM$ BHJ solar cells characterized by IMVS demonstrated that post-annealing reduced the recombination of electron carriers in the device. The recombination times of $P3HT/PC_{61}BM$ BHJ solar cells post-annealed at room temperature, 80, 120, and $140^{\circ}C$ were 0.009, 0.020, 0.024, and 0.030 ms, respectively, at a short-circuit current of 0.18 mA. The results indicated that the IMVS analysis can be effectively used as powerful.

Phylogenetic and Recombination Analysis of Apple Stem Grooving Virus Isolates from Pears in Korea

  • Nam-Yeon Kim;Rae-Dong Jeong
    • Research in Plant Disease
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    • v.29 no.2
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    • pp.193-199
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    • 2023
  • The apple stem grooving virus (ASGV) is one of the most harmful latent viruses infecting pear orchards worldwide. To examine the genetic diversity of ASGV in Korean pear orchards, the complete coat protein (CP) gene of five ASGV isolates collected from various regions were identified. The five Korean ASGV isolates showed 88-96% nucleotide identity with the 11 isolates worldwide occurring elsewhere in the world. Phylogenetic analysis of five isolates, as well as the previously sequenced isolates, indicated that the ASGV clusters had no correlation with the host or geographical regions of origin. Recombination analysis showed that one of the five Korean isolates is a recombinant, with a recombination site in the CP gene region (nt 532-708). This study is the first report of natural recombination within the CP gene of ASGV isolates from pears grown in Korea.

A Yeast MRE3/REC114 Gene is Essential for Normal Cell Growth and Meiotic Recombination

  • Leem, Sun-Hee
    • Journal of Microbiology
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    • v.37 no.4
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    • pp.248-255
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    • 1999
  • We have analyzed the MRE3/REC114 gene of Saccharomyces cerevisiae, previously detected in isolation of mutants defective in meiotic recombination. We cloned the MRE3/REC114 gene by complementation of the meiotic recombination defect and it has been mapped to chormosome XIII. The DNA sequence analysis revealed that the MRE3 gene is identical to the REC114 gene. The upstream region of the MRE3/REC114 gene contains a T_4C site, a URS (upstream repression sequence) and a TR (T-rich) box-like sequence, which reside upstream of many meiotic genes. Coincidentally, northern blot analysis indicated that the three sizes of MRE3/REC114 transcripts, 3.4, 1.4 and 1.2 kb, are induced in meiosis. A less abundant transcript of 1.4 kb is detected in both mitotic and meiotic cells, suggesting that it is needed in mitosis as well as meiosis. To examine the role of the MRE3/REC114 gene, we constructed mre3 disruption mutants. Strains carrying an insertion or null deletion of the MRE3/REC114 gene showed slow growth in nutrient medium and the doubling time of these cells increased approximately by 2-fond compared to the wild-type strain. Moreover, the deletion mutant (${\delta}$mre3) displayed no meiotically induced recombination and no viable spores. The mre3/rec114 spore lethality can be suppressed by spo13, a mutation that causes cells to bypass reductional division. The double-stranded breaks (DSBs) which are involved in initiation of meiotic recombination were not detected in the analysis of meiotic chromosomal DNA from the mre3/rec114 disruptant. From these results we suggest that the MRE3/REC114 gene product is essential in normal growth and in early meiotic stages involved in meiotic recombination.

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