• Title/Summary/Keyword: repeat region

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Stability of Human Centromeric Alphoid DNA Repeat during Propagation in Recombination-Deficient Yeast Strains (효모의 재조합 변이주를 이용한 인간 Centromeric Alphoid DNA Repeat의 안정성에 관한 연구)

  • Kim, Kwang-Sup;Shin, Young-Sun;Lee, Sang-Yeop;Ahn, Eun-Kyung;Do, Eun-Ju;Park, In-Ho;Leem, Sun-Hee;SunWoo, Yang-Il
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.243-249
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    • 2007
  • The centromere is a highly differentiated structure of the chromosome that fulfills a multitude of essential mitotic and meiotic functions. Alphoid DNA (${\alpha}$-satellite) is the most abundant family of repeated DNA found at the centromere of all human chromosomes, and chromosomes of primates in general. The most important parts in the development of Human Artificial Chromosomes (HACs), are the isolation and maintenance of stability of centromeric region. For isolation of this region, we could use the targeting hook with alphoid DNA repeat and cloned by Transformation-Associated Recombination (TAR) cloning technique in yeast Saccharomyces cerevisiae. The method includes rolling-circle amplification (RCA) of repeats in vitro to 5 kb-length and elongation of the RCA products by homologous recombination in yeast. Four types of $35\;kb{\sim}50\;kb$ of centromeric DNA repeat arrays (2, 4, 5, 6 mer) are used to examine the stability of repeats in homologous recombination mutant strains (rad51, rad52, and rad54). Following the transformation into wild type, rad51 and rad54 mutant strains, there were frequent changes in inserted size. A rad52 mutant strain showed extremely low transformation frequency, but increased stability of centromeric DNA repeat arrays at least 3 times higher than other strains. Based on these results, the incidence of large mutations could be reduced using a rad52 mutant strain in maintenance of centromeric DNA repeat arrays. This genetic method may use more general application in the maintenance of tandem repeats in construction of HAC.

Induction of RNA-mediated Resistance to Papaya Ringspot Virus Type W

  • Krubphachaya, Pongrit;Juricek, Mila;Kertbundit, Sunee
    • BMB Reports
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    • v.40 no.3
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    • pp.404-411
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    • 2007
  • Transformation of cantaloupes with the coat protein (cp) gene of papaya ringspot virus type W (PRSV-W), Thai isolate, was used to introduce virus resistance. Binary vectors containing either the full length coat protein coding region under control of the 35S CaMV promoter(pSA1175), or the inverted-repeat of a coat protein coding region (pSA1304), were constructed and used for Agrobacteriummediated transformation of cotyledonary explants of the cantaloupe cultivar Sun Lady. Four independent transgenic lines were obtained using pSA1304 and one using pSA1175. Integration of the PRSV-W cp gene into the genome of these transgenic lines was verified by PCR amplification, GUS assays and Southern blot hybridization. In vitro inoculation of these lines with PRSV-W revealed that whereas the line containing pSA1175 remained sensitive, the four lines containing pSA1304 were resistant. The presence of small RNA species, presumably siRNA, corresponding to regions of the viral cp gene in transgenic lines resistant to PRSV-W supports the involvement of post-transcriptional gene silencing in the establishment of resistance.

Investigation of Deletion Variation and Methylation Patterns in the 5' LTR of Porcine Endogenous Retroviruses

  • Jung, K.C.;Simond, D.M.;Moran, C.;Hawthorne, W.J.;Jeon, J.T.;Jin, D.I.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.11
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    • pp.1572-1575
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    • 2008
  • The xenotransplantation of pig organs and cells can be related with a risk of transmission of infectious diseases to human. Previous findings indicate that the regulatory region of PERV for retroviral transcription, replication and integration into the cellular DNA is located on the 5' Long Terminal Repeat (LTR). The objective of this study is the investigation of methylation and deletion status of the PERV 5' LTR region which can be used for regulating PERV expression. We compared the sequences of genomic DNA and bisulfite-treated genomic DNA from PK-15 cells expressing PERV to observe the methylation status of the 5' LTR. Our results showed that the CpG sites of U3 were methylated and methylation was inconsistent in the R and U5 regions. Also, variable numbers of 18 bp repeats and 21 bp repeats were detected on 5' LTR by sequencing analysis. The consistent U3 methylation might be indicative of host suppression of expression of the retroviruses.

Wear of UHMWPE Pins Against Ti-alloy and Stainless Steel Disks Moving in Two Kinematic Motions (두가지 기구운동을 하는 타이타늄 합금과 스테인레스 스틸 디스크에 대한 초고분자량 폴리에틸렌 핀의 마멸)

  • 이권용;김석영;김신윤
    • Tribology and Lubricants
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    • v.18 no.2
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    • pp.167-172
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    • 2002
  • The wear behaviors of ultrahigh molecular weight polyethylene pins against titanium alloy and stainless steel disks moving in two different kinematic motion were investigated by conducting repeat pass rotational sliding and linear reciprocal sliding wear tests. Linear reciprocal motion wore more the polyethylene pin than did repeat pass rotational motion for both disk materials. It means that the repeated directional change of contact stresses generates more wear debris in polyethylene. For the linear reciprocal sliding tests, titanium alloy disks were damaged with some scratches after one million cycles but no surface damage was observed on the polyethylene pins. On the other hand, fur the repeat pass rotational sliding tests, all titanium alloy disks were severely abraded on the entire region of sliding track. This phenomenon can be interpreted by that stress fatigue under repeated sliding contact initiated titanium oxide layer wear particles from disk surface, and these hard particles were embedded into polyethylene pin and then they severely abraded the disk surface. From these results it can be concluded that the kinematic motion in pin-on-disk wear tests play a crucial role on the wear behaviors of UHMWPE pins against titanium alloy and stainless steef discs.

Association of the X-linked Androgen Receptor Leu57Gln Polymorphism with Monomelic Amyotrophy

  • Park, Young-Mi;Lim, Young-Min;Kim, Dae-Seong;Lee, Jong-Keuk;Kim, Kwang-Kuk
    • Genomics & Informatics
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    • v.9 no.2
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    • pp.64-68
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    • 2011
  • Monomelic amyotrophy (MA), also known as Hirayama disease, occurs mainly in young men and manifests as weakness and wasting of the muscles of the distal upper limbs. Here, we sought to identify a genetic basis for MA. Given the predominance of MA in males, we focused on candidate neurological disease genes located on the X chromosome, selecting two X-linked candidate genes, androgen receptor (AR ) and ubiquitin-like modifier activating enzyme 1 (UBA1). Screening for genetic variants using patients' genomic DNA revealed three known genetic variants in the coding region of the AR gene: one nonsynonymous single-nucleotide polymorphism (SNP; rs78686797) encoding Leu57Gln, and two variants of polymorphic trinucleotide repeat segments that encode polyglutamine (CAG repeat; rs5902610) and polyglycine (GGC repeat; rs3138869) tracts. Notably, the Leu57Gln polymorphism was found in two patients with MA from 24 MA patients, whereas no variants were found in 142 healthy male controls. However, the numbers of CAG and GGC repeats in the AR gene were within the normal range. These data suggest that the Leu57Gln polymorphism encoded by the X-linked AR gene may contribute to the development of MA.

Wear of UHMWPE Pins against Ti-alloy and Stainless Steel Disks Moving in Two Kinematic Motions (두가지 기구운동을 하는 타이타늄 합금과 스테인레스 스틸 디스크에 대한 초고분자량 폴리에틸렌 핀의 마멸)

  • 이권용;김석영;김신윤
    • Proceedings of the Korean Society of Tribologists and Lubrication Engineers Conference
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    • 2000.11a
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    • pp.67-71
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    • 2000
  • The wear behaviors of ultrahigh molecular weight polyethylene pins against titanium alloy and stainless steel disks moving in two different kinematic motion were investigated by conducting repeat pass rotational sliding and linear reciprocal sliding wear tests. Linear reciprocal motion wore more the polyethylene pin than did repeat pass rotational motion for both disk materials. It means that the repeated directional change of contact stresses generates more wear debris in polyethylene. For the linear reciprocal sliding tests, titanium alloy disks were damaged with some scratches after one million cycles but no surface damage was observed on the polyethylene pins. On the other hand, for the repeat pass rotational sliding tests, all titanium alloy disks were severely abraded on the entire region of sliding track. This phenomenon can be interpreted by that stress fatigue under repeated sliding contact initiated titanium oxide layer wear particles from disk surface, and these hard particles were embedded into polyethylene pin and then they severely abraded the disk surface. From these results it can be concluded that the kinematic motion in pin-on-disk wear tests play a crucial role on the wear behaviors of UHMWPE pins against titanium alloy and stainless steel disks.

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Genetic Mapping of a Resistance Locus to Phytophthora sojae in the Korean Soybean Cultivar Daewon

  • Jang, Ik-Hyun;Kang, In Jeong;Kim, Ji-Min;Kang, Sung-Taeg;Jang, Young Eun;Lee, Sungwoo
    • The Plant Pathology Journal
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    • v.36 no.6
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    • pp.591-599
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    • 2020
  • Phytophthora root and stem rot reduce soybean yields worldwide. The use of R-gene type resistance is currently crucial for protecting soybean production. The present study aimed to identify the genomic location of a gene conferring resistance to Phytophthora sojae isolate 2457 in the recombinant inbred line population developed by a cross of Daepung × Daewon. Singlemarker analysis identified 20 single nucleotide polymorphisms associated with resistance to the P. sojae isolate 2457, which explained ~67% of phenotypic variance. Daewon contributed a resistance allele for the locus. This region is a well-known location for Rps1 and Rps7. The present study is the first, however, to identify an Rps gene locus from a major soybean variety cultivated in South Korea. Linkage analysis also identified a 573 kb region on chromosome 3 with high significance (logarithm of odds = 13.7). This genomic region was not further narrowed down due to lack of recombinants within the interval. Based on the latest soybean genome, ten leucine-rich repeat coding genes and four serine/ threonine protein kinase-coding genes are annotated in this region, which all are well-known types of genes for conferring disease resistance in crops. These genes would be candidates for molecular characterization of the resistance in further studies. The identified R-gene locus would be useful in developing P. sojae resistant varieties in the future. The results of the present study provide foundational knowledge for researchers who are interested in soybean-P. sojae interaction.

Analysis for the function of the core region of Bacillus polymyxa CFTase

  • Kwon, Hyun-Ju;You, Kyung-Ok;Park, Ju-Hee;Oh, You-Na;Kim, Kwang-Hyun;Kim, Byung-Woo
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.582-585
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    • 2003
  • Sequence analysis indicated that Bacillus polymyxa MGL21 CFTase was divided into five distinct regions. CFTase contained three regions of repeat sequences at the N-terminus and C-terminus. The endo-inulinase region of homology (ERH) of CFTase was similar to that of Pseudomonas mucidolens endo-inulinase. Furthemore, CFTase possessed a highly conserved core region. In order to understand the role of the core region on the function of CFTase from B. polymyxa MGL21 CFTase ${\Delta}NC$ was prepared. The molecular weight of the purified wild type CFTase and $CFTase{\Delta}NC$ were 148kDa, 90kDa, respectively.

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Tandem Repeats (CCTTT)n in the Promoter of iNOS Gene in Korean Genome

  • Baek, Sun-Ah;Yoo, Min
    • Biomedical Science Letters
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    • v.15 no.2
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    • pp.167-170
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    • 2009
  • Nitric oxide is an important factor to regulate the biochemical reactions in the body such as expansion of blood vessel, neural conduction and antimicrobial activity. There are two forms of nitric oxide synthase and iNOS has attracted most attention because it is involved in the development of diabetes and cardiac disease condition. There are several regulatory sequences in the promoter region of iNOS gene. One of them is (CCTTT)n. It has been reported that the number of tandem repeat of (CCTTT)n varies from population to population. So, we analyzed (CCTTT)n polymorphism in Korean genome for the purpose of comparison. According to our present study Koreans are different from other Asians reported previously because $(CCTTT)_{10}$ is the highest incidence as opposed to $(CCTTT)_{12}$ for other countries. This study should facilitate the understanding of the expression of iNOS gene in different population.

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Relationship between Oxidation and Wear of Ultra-High Molecular Weight Polyethylene for Total Joint Arthroplasty

  • Lee, Kwon-Yong
    • KSTLE International Journal
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    • v.2 no.1
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    • pp.55-58
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    • 2001
  • The most widely-used orthopaedic grade polymer bearing liner material, ultrahigh molecular weight polyethylene (UHMWPE), for the total joint arthroplasty degrades after gamma-irradiation sterilization through the progressive oxidation in a shelf and in vivo. Oxidative degradation makes UHMWPE brittle and leads to decrease in mechanical properties. In this study the relationship between post-gamma-irradiation aging time and wear of UHMWPE was investigated. Six retrieved polyethylene hip liners implanted for 3-16 years and then stored in air for 1.5-6.5 years until tests were used. Two types of pin-on-disk wear testing were conducted by the uni-directional repeat pass rotating and by the linear reciprocating stainless steel disks against stationary polyethylene pins under 4Mpa at 1Hz with bovine serum lubrication in ambient environment. Wear of retrieved polyethylene hip liners does not have direct correlation with in vivo or total aging time. Linear reciprocal sliding motion generated more remarkable wear than uni-directional repeat pass sliding motion. It indicates that kinematic motion affects very crucially on the wear of aged UHMWPE having brittle white band region.

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