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Restriction endonuclease analysis of mitochondrial DNA of Acanthamoebn sp. YM-4 (Korean isolate) (Acanrhamoeba sp. YM-4의 미토콘드리아 DNA의 RFLP분석)

  • Sin, Ho-Jun;Im, Gyeong-Il;Jeon, Gwang-U
    • Parasites, Hosts and Diseases
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    • v.35 no.2
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    • pp.119-126
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    • 1997
  • Acanthnmoebn sp. YM-4 is simitar to A. culbertsoni based upon morphological characteristics of trophozoites and cysts. However, based on other characteristics, pathogenicity to mice, in uitro cytotoxicity and isoenzyme patterns, Acanthomoebo sp. YM- 4 was quite different from A. culbertsoni. Restriction fragment length polymorphism (RFLP) analysis of mtDNA is useful in the classification of members belonging to the genus Acanthcmoebn. Therefore, in this study, RFLP analysis of Acnnthcmoeba mtDNAs was accomplished using five restriction enzymes: Hnelll, Hinull, Clcl, Pudl and ScE. Each restriction enzyme produced approximately 3-15 fragments (range: from 0:6 kip to 34.4 kbp) . The mtDNA genome size, calculated by the summation of restriction fragments, averaged 46.4 kbp in Acnnthamoeba sp. YM-4,48.3 kbp in A. culbertsoni and 48.8 kbp in A. polyphaic, respectively. Digested mtDNA fragments of Accnthcmoeba sp. YM-4 contained nine and seven same size fragments, respectively, from a total of 67 and 69 fragments observed in A. culbertsoni and A. polyphcgn. An estimate of the genetic divergence was 10.1% between Acanthamoebc sp. YM-4 and A. culbertsoni, and 9.9% between Acanthamoebn sp. YM-4 and A. polyphcga.

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On the wing venation and scales of Dendrolimus spectabilis Butler (I) (솔나방의 시맥(翅脈)과 인편(鱗片)에 관(關)한 연구(硏究) (I))

  • Yun, Jeong Koo
    • Journal of Korean Society of Forest Science
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    • v.2 no.1
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    • pp.59-65
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    • 1962
  • The objects of this experiment are to find out the local variation of the Dendrolimus Spectabilis Butler, of which sample was first collecteted 15 bodies of male and 35 bodies of female adult at Suwon area. the wing veins and the scale shape have been observed through the microscope (100) and the scale size (from the bottom of the scale to the top of the lobe) has also been measured by the micrometer. The results of this experiment are as follows: 1. There is nodifference between the venation of the male body and that of the female body. Also we can not find any differences between the right and the left wing, and between each body. The fore wings consit of 13 longitudinal veins and the only one "V" shape cross vein which is between the 5th and 6th vein. The hind wings consist of 9 longitudinal veins and the only one "V" shape cross vein which is mentioned above. 2. The scale types are divided into 4 Groups in its shape. (A) The scales of I Group are short and the lower parts of them almost look like a circle, having a small projection at their bottom. The upper parts of them have 2 or 10 lobes. We can find the lobes with fine hairs or the lobes without them at the top of the scales. (B) The scales of II Group are longer than that of I Group. The shape of the lower parts of the scales is similar to that of I Group. The upper parts of the scales have 2 or 10 lobes. (C) The scales of III Group are long and almost alike in a long wedgeshape. The upper parts of the scales have 2 Or IO lobes and we can find long fine hairs at the top of each lobe. (D) The scales of IV Group are long and the shape of the lower parts of the scales is similar to that of III Group. The lobes are short and not sharp. We can find 2 or 9 lobes. 3. The scales of I Group and II Group are distributed on the whole surface except on the outer margin. The most scales of III Group are distributed on the wing base. The scales of IV Group are distributed on the outer margin only. The scales with 4 or 5 lobes are most widely distributed not considering their Group or distributing parts. 4. In I Group the variation of the scale length become gradually greater as the number of the lobes are increasing. In II, III, IV Group, however, the variation of the scale length is in direct opposition to the above mentioned. The variation of the scale width becomes pretty small in any Groups and the scale width become wider as the number of the lobes are increasing. 5. The source of the wing colouration is pigmnetal colour, and the wing colouration is expressed by the amount of brown and white scales.

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Genome Type Analysis of Adenovirus Serotypes 1, 2 and 5 Isolated from Children with Lower Respiratory Tract Infections in Korea (하기도 감염 환아에서 분리된 Adenovirus 1, 2, 5 혈청형의 유전체형 분석)

  • Park, Ki-Won;Choi, Eun-Hwa;Choun, Ji-Tae;Lee, Hoan-Jong;Park, Ki-Ho
    • Pediatric Infection and Vaccine
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    • v.12 no.2
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    • pp.166-177
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    • 2005
  • Purpose : The purpose of this study was to examine the molecular epidemiology and genetic variability of adenovirus(Ad) serotypes Ad1, Ad2, and Ad5 over 14 years in Korea. Methods : A total of 382 adenoviral strains isolated from the nasopharyngeal aspirates of children with lower respiratory tract infections in Seoul, Korea from November 1990 to February 2003 were serotyped by neutralization assay with type-specific antisera. Viral DNAs were extracted from infected cell lysates by the modified Hirt procedure. Genome type(GT) was determined by DNA restriction analysis with 12 restriction enzymess(BamHI, BclI, BglI, BglII, BstEII, EcoRI, HindIII, HpaI, SalI, SmaI, XbaI, and XhoI). To evaluate the genetic relatedness, pairwise comigrating restriction fragments(PCRF) analysis was performed. Results : Of 382 strains, 33 strains(9%) were Ad1, 45 strains(12%) were Ad2, and 24 strains(6%) were Ad5. Eighteen GTs(Ad1p1-Ad1p7, Ad1a, Ad1b, Ad1b1-Ad1b3, Ad1c, Ad1d, Ad1e, Ad1e1, Ad1e2, Ad1f) among Ad1, 24(Ad2p1-Ad2p11, Ad2a, Ad2a1-Ad2a6, Ad2b, Ad2c, Ad2d, Ad2e, Ad2e1-Ad2e3) among Ad2, and 10(Ad5p1, Ad5p2, Ad5a, Ad5a1-Ad5a7) among Ad5 strains were identified. One or two strains of the vast majority of GTs were isolated during the study period while a few GTs were identified sporadically with more than 2 strains. It is notable that some GTs such as Ad1p5 and Ad5a1 appeared in cluster during a short period. In analysis of genetic relatedness, the degree of PCRFs(pairwise comigrating restriction fragments) for Ad1 varied from 79 to 99%, for Ad2, 82 to 99%, and for Ad5, 85 to 99%. Conclusion : This study established the comprehensive nomenclature systems of Ad1, Ad2, and Ad5. Diverse GTs identified in this study have crucial implications in the genomic diversity and epidemiological characteristics of Ad1, Ad2, and Ad5.

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Screening and Characterization of LTR Retrotransposons in the genomic DNA of Pleurotus eryngii (큰느타리버섯 유전체내 LTR Retrotransposon 유전자 탐색 및 특성연구)

  • Kim, Sinil;Le, Quy Vang;Kim, Sun-Mi;Ro, Hyeon-Su
    • The Korean Journal of Mycology
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    • v.42 no.1
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    • pp.50-56
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    • 2014
  • Transposable elements (TEs) are mobile DNA elements that often cause mutations in genes and alterations in the chromosome structure. In order to identify and characterize transposable elements (TEs) in Pleurotus eryngii, a TE-enriched library was constructed using two sets of TE-specific degenerated primers, which target conserved sequences of RT and RVE domains in fungal LTR retrotransposons. A total of 256 clones were randomly chosen from the library and their insert sequences were determined. Comparative investigation of the insert sequences with those in repeat element database, Repbase, revealed that 71 of them were found to be TE-related fragments with significant similarity to LTR retrotransposons from other species. Among the TE sequences, the 70 TEs were Gypsy-type LTR retrotransposons, including 20 of MarY1 from Tricholoma matsutake, 26 of Gypsy-8_SLL from Serpula lacrymans, and 16 of RMER17D_MM from mouse, whereas a single sequence, Copia-48-PTR, was found as only Copia-type LTR retrotransposon. Southern blot analysis of the HindIII-digested P. eryngii genomic DNA showed that the retrotransposon sequences similar to MarY1 and Gypsy-8_SLL were contained as high as 14 and 18 copies per genome, respectively, whereas other retrotransposons were remained low. Moreover, both of the two Gypsy retrotransposons were expressed in full length mRNA as shown by Northern blot analysis, suggesting that they were functionally active retrotransposons.

Mapping of RFLP Markers Linked to Bacterial Blight Resistant Genes (Xa-1, Xa-3) in Rice (벼 흰잎마름병 저항성 유전자(Xa-1, Xa-3)연관 RFLP 마커 탐색)

  • 강현중;김현순;남정권;이영태;이승엽;김석동
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.48 no.6
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    • pp.419-423
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    • 2003
  • Bacterial blight caused by Xantomonas oryzae pv. oryzae is one of the most serious diseases of rice especially in southern area of Korea. Three races, $\textrm{K}_1$, $\textrm{K}_2$ and $\textrm{K}_3$, are the most dominant species. lo improve rice breeding efficiency using marker assisted selection, some RFLP markers were surveyed for polymorphism between resistant and susceptible to $\textrm{K}_1$ and $\textrm{K}_3$. And, 127 doubled-haploid (DH) lines derived from Milyang121/HRl1650-1-4-2 and 131 DH lines derived from Milyang123/HR10624-AC5 were evaluated to bacterial blight ($\textrm{K}_1$ and $\textrm{K}_3$). Milyang121 and HR10624-AC5 have Xa-1, resistant to $\textrm{K}_1$ race, and Milyang123 has Xa-3, resistant to $\textrm{K}_1$ and $\textrm{K}_3$ race. Three markers, RZ590, RZ536 and RG303, showing polymorphism between parents and resistance gene, Xa-1 and Xa-3, were analysed in the two combinations of DH lines. The segregation pattern of resistant DH population of Milyang123/HR10624-AC5 to susceptible showed 3:1 and 1:1 in $\textrm{K}_1$ and $\textrm{K}_3$ race. In three RFLP markers, RZ590 was linked to Xa-1 on chromosome 4, and RZ536 and RG303 were linked to Xa-3 on chromosome 11. The map distance between Xa-1 and RZ590 was 3.1cM on chromosome 4, and Xa-3 and RZ536/RG303 were 7.6/16.0cM on chromosome 11, respectively. The results of RFLP mapping will be useful for the selection and pyramiding of bacterial blight resistant genes.

Characterization of a Novel cry1-Type Gene from Bacillus thuringiensis subsp. alesti Strain LY-99

  • Qi, Xu Feng;Li, Ming Shun;Choi, Jae-Young;Roh, Jong-Yul;Song, Ji Zhen;Wang, Yong;Jin, Byung-Rae;Je, Yeon-Ho;Li, Jian Hong
    • International Journal of Industrial Entomology and Biomaterials
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    • v.18 no.1
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    • pp.18-27
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    • 2009
  • B. thuringiensis strain LY-99 belonging to subsp. alesti (H3a3c), was isolated from Chinese tobacco warehouse and showed significantly high toxicity to Plutella xylostella. For the identification of the cry1-type genes from B. thuringiensis LY-99, an extended multiplex PCRrestriction fragment length polymorphism (PCRRFLP) method was established by using two pairs of universal primers based on the conserved regions of the cry1-type genes to amplify around 2.4 kb cry1-type gene fragments. Then the DNA fragment was cloned into pGEM-T Easy vector and digested with EcoRI and EcoRV enzymes. Through this method, a known cry1-type gene was successfully identified from the reference strain, B. thuringiensis subsp. alesti. In addition, the RFLP patterns revealed that B. thuringiensis LY-99 included a novel cry1A-type gene in addition to cry1Aa, cry1Ac, cry1Be and cry1Ea genes. The novel cry1A-type gene was designated cry1Ah2 (Genbank accession No DQ269474). An inverse PCR method was used to amplify the flank regions of cry1Ah2 gene. Finally, 3143 bp HindIII fragment from B. thuringiensis LY-99 plasmid DNA including 5' region and partial ORF was amplified, and sequence analysis revealed that cry1Ah2 gene from LY-99 showed 89.31% of maximum sequence similarity with cry1Ac1 crystal protein gene. In addition, the deduced amino acid sequence of Cry1Ah2 protein shared 87.80% of maximum identity with that of Cry1Ac2. This protein therefore belongs to a new class of B. thuringiensis crystal proteins.

Analysis of nucleotide sequence of a novel plasmid, pILR091, from Lactobacillus reuteri L09 isolated from pig

  • Lee, Deog-Yong;Kang, Sang-Gyun;Rayamajhi, Nabin;Kang, Milan;Yoo, Han Sang
    • Korean Journal of Veterinary Research
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    • v.48 no.4
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    • pp.441-449
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    • 2008
  • The genus Lactobacillus is the largest of the genera included in lactic acid bacteria and is associated with mucosal membranes of human and animal. Only a few Lactobacillus plasmid-encoded functions have been discovered and used. In this study, a novel plasmid (pILR091) was isolated from a wild L. reuteri isolated from pig and described the characteristics of its replicons, genetic organization, and relationship with other plasmids. After digestion of the plasmid, pILR091, with SalI, plasmid DNA was cloned into the pQE-30Xa vector and sequenced. The complete sequence was confirmed by the sequencing of PCR products and analyzed with the Genbank database. The isolate copy number and stability were determined by quantitative-PCR. The complete sequence of L. reuteri contained 7,185 nucleotides with 39% G-C content and one cut site by two enzymes, SalI and HindIII. The similar ori sequence of the pC194- rolling circle replication family (TTTATATTGAT) was located 63 bp upstream of the protein replication sequence, ORF 1. Total of five ORFs was identified and the coding sequence represented 4,966 nucleotides (70.4%). ORF1 of pILR091 had a low similarity with the sequence of pTE44. Other ORFs also showed low homology and E-values. The average G-C content of pILR091 was 39%, similar with that of genomic DNA. The copy number of pILR091 was determined at approximately 24 to 25 molecules per genomic DNA. These results suggested that pILR091 might be a good candidate to construct a new vector, which could be used for cloning and expression of foreign genes in lactobacilli.

Helicobacter pylori Strain 51 (Korean Isolate): Ordered Overlapping BAC Library, Combined Physical and Genetic Map, and Comparative Analysis with H. pylori Strain 26695 and Strain J99

  • KANG HYUNG-LYUN;LEE WOO-KON;SONG JAE-YOUNG;CHOI SANG-HAENG;PARK SEONG-GYU;RYU BOK-DEOK;LEE EUN-JOO;KIM JI-SUN;PARK JEONG-UCK;BAIK SEUNG-CHUL;CHOI MYOUNG-BUM;YOUN HEE-SHANG;KO GYUNG-HYUCK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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    • pp.844-854
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    • 2005
  • We constructed a defined physical and genetic map of H. pylori strain 51, previously isolated from a Korean patient with a duodenal ulcer, by combining a restriction analysis by pulse-field gel electrophoresis with the construction of a BAC library. A Notl-digest of H. pylori strain 51 genome yielded seven fragments, from which the genomic size was estimated to be 1,698$\pm$24 kb. The BAC library was constructed from 50 to 200 kb fragments of HindIII-digested genomic DNA. From 700 BAC clones, an ordered overlapping maxi-set of 82 BAC clones was assembled that covered the entire genome. The positions of 15 genes were localized in the strain 51 genome with 4-22 kb of resolution and were compared with their orthologues in strain 26695 and strain J99. The arrangement of the 15 genes was identical in strain 51 and strain J99, except for flaA and hpaA. The plasticity zone of strain 51, like that of strain J99, was located in the single region, and was shorter than those of strain 26695 and strain J99. The strain 51 plasticity zone consisted of ORFs common only to strain 51 and J99 or to strain 51 and 26695, as well as strain 51-specific ORFs. Three genetic translocations and/or inversions were found between orthologue ORFs in strain 51 and strain J99. These results show that the chromosomal organization of strain 51 differs from Western strains such as strain 26695 and strain J99.

Transcriptional regulation of soybean ${\beta}-conglycinin$ gene expression. -(I) Identification of a soybean embryo factor interacting with upstream region of soybean ${\beta}-conglycinin$ gene- (대두 ${\beta}-conglycinin$ 유전자 발현의 전사 조절에 관한 연구 -(I) 대두 ${\beta}-conglycinin$ 유전자의 upstream 영역에 결합하는 대두 배 인자의 동정-)

  • Lee, Jeong-Yeon;Chung, Dong-Hyo;Kim, Woo-Yeon
    • Applied Biological Chemistry
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    • v.36 no.6
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    • pp.547-552
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    • 1993
  • Soybean nuclear extracts and S-100 were prepared to examine the soybean embryo factors which bind to the upstream region of soybean ${\beta}-conglycinin$ ${\alpha}'$ subunit gene. SEF3(soybean embryo factor 3), which is presumed to be a trans-acting factor for the expression of the gene, was detected in gel mobility shift assay using the DNA probe containing two AACCCA hexanucleotides. DNA probe containing CATGCAT or AACACA was used to find any other soybean embryo factor interacting with the upstream region of ${\beta}-Conglycinin$ ${\alpha}'$ subunit gene. It was found that there was no common DNA binding protein detected both in nuclear extracts and S-100. The relative levels of SEF3 binding activity both in nuclear extracts and S-100 of maturing soybean seeds were determined. SEF3 activity of nuclear extracts was first detected around 20 days after pollination and significantly increased around 32 days after pollination.

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Cell Surface Display of Arylsulfatase Gene from Pseudoalteromonas carageenovora in Saccharomyces cerevisiae (Saccharomyces cerevisiae에서 Pseudoalteromonas carageenovora 유래 Arylsulfatase 유전자의 표층 발현)

  • Cho, Eun-Soo;Kim, Hyun-Jin;Jung, So-A;Kim, Jeong-Hwan;Kim, Yeon-Hee;Nam, Soo-Wan
    • Microbiology and Biotechnology Letters
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    • v.37 no.4
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    • pp.355-360
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    • 2009
  • In this study, the arylsulfatase gene (astA, 984 bp ORF) from Pseudoalteromonas carrageenovora genome was expressed on the cell surface of S. cerevisiae by fusing with Aga2p linked to the membrane anchored protein, Aga1p. The constructed plasmid, pCTAST (7.1 kb), was introduced to S. cerevisiae EBY100 cell, and yeast transformants on YPDG plate showed the hydrolyzing activity for 4-methylumbelliferyl-sulfate and p-nitrophenyl-sulfate. When S. cerevisiae EBY100/pCTAST was grown on YPDG medium, the arylsulfatase activity of cell pellet reached about 1.2 unit/mL, whereas no extracellular arylsulfatase activity was detected. The DNA ladder in agarose prepared from agar by this recombinant arylsulfatase showed similar resolution and migration patterns with a commercial agarose. This results revealed that arylsulfatase expressed on the cell surface of S. cerevisiae could be applicable to the economic production of electrophoretic-grade agarose.