• 제목/요약/키워드: mtDNA RFLP

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Morphological and Molecular Classifications of Genus Pholis

  • Lee, Sung-Hoon;Jang, Yo-Soon;Baik, Chung-Boo;Han, Kyeong-Ho;Myung, Jung-Goo;Lee, Jin-Hee;Choi, Sang-Duk;Kim, Seon-Jae;Kim, Jong-Oh;Hwang, Jae-Ho
    • Animal cells and systems
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    • 제13권4호
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    • pp.453-460
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    • 2009
  • Morphological and molecular classifications were attempted in an effort to establish species-specific classifications of three species of the genus Pholis in Korea; these species were subjected to morphological and molecular methodologies using body measurements, RFLP, RAPD, and phylogenetic trees using the nucleotide sequences of mitochondrial 16S and 12S ribosomal DNAs, cytochrome c oxidase I, and cytochrome b. The data demonstrated that the three species of genus Pholis are distinct from each other, both morphologically and genetically.

불임여성에서 NAT2, GSTM1, CYP1A1 유전자 다형성과 자궁내막증의 상관관계에 관한 연구 (Association between Endometriosis and Polymorphisms of N-acetyl Transferase 2 (NAT2), Glutathione S-transferase M1 (GSTM1) and Cytochrome P450 (CYP) 1A1 Genes in Korean Infertile Patients)

  • 송현정;전진현;최혜원;허걸;강인수;궁미경;이형송
    • Clinical and Experimental Reproductive Medicine
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    • 제31권2호
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    • pp.141-147
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    • 2004
  • Objective: To investigate the association between endometriosis and polymorphisms of N-acetyl transferase 2 (NAT2), glutathione S-transferase M1 (GSTM1), and cytochrome P450 (CYP) 1A1 genes in Korean infertile patients. Materials and Methods: A total of 303 infertile patients who had undertaken diagnostic laparoscopy during January, 2001 through December, 2003 at Samsung Cheil Hospital enrolled in this study. The patients were grouped according to laparoscopic findings: minimal to mild endometriosis (group I: n=147), moderate to severe endometriosis (group II: n=57), normal pelvic cavity (n=99). Peripheral blood was obtained and genomic DNA was extracted. The genotypes of each genes were analyzed using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). For NAT2, RFLP was used to detect the wild type (wt) and mutant (mt) alleles, enabling classification into slow (mt/mt) or fast (wt/wt or wt/mt) acetylation genotypes. For GSTM1, PCR was used to distinguish active (+/- or +/+) from null (-/-) genotypes. For CYP1A1, MspI digestion was used to detect the wild type (A1A1), heterozygote (A1A2) or mutant (A2A2) genotypes. Result: The genotype frequencies of NAT2 slow acetylator was 12.8%, 10.9%, 12.8% in group I, group II and control, respectively. The genotype frequencies of GSTM1 null mutation was 55.3%, 41.8%, 53.2% in group I, group II and control, respectively. The genotype frequencies of CYP1A1 MspI polymorphism was 16.3%, 9.1%, 18.1% in group I, group II and control, respectively. No significant difference was observed between endometriosis and normal controls in the genotype frequencies of the NAT2, GSTM1, CYP1A1 MspI polymorphism. Conclusion: The NAT2, GSTM1, CYP1A1 gene polymorphism may not be associated with the susceptibility of endometriosis in Korean women.

미토콘드리아 DNA의 제한효소 분석법에 의한 영지의 계통분류 (Phylogeny of Ganoderma Based on the Restriction Enzyme Analysis of Mitochondrial DNA)

  • 홍순규;정학성
    • 미생물학회지
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    • 제32권4호
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    • pp.245-251
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    • 1994
  • 영지속(Ganoderma)에 속하는 7종 10균주에 대하여 미토콘드리아 DNA의 제한효소 분절양상 비교를 통한 계통분석을 수행하였다. 여러 가지 제한효소들 중 생산된 절편이 충분한 정보를 가지고 있으면서 서로 구별할 수 있는 6가지의 제한효소를 분석에 이용하였다. 절편양상을 설 비교하여 전체 절편중 공통된 절편의 개수를 구하고 이로부터 염기위치당 염기치환율을 구하였으며, 이를 균주간의 진화거리로 계산하여 PHYLIP package의 Neighbor-joining 방법에 이한 계통도를 얻고 그 결과를 고찰하였다. 특이한점은 G. lucidum의 3균주와 G. lobatum 이 유연관계가 많이 있다는 점이다. 이러한 결과는 G. lucidum과 G. lobatum은 종의 다양성으로 인하여 과거부터 복합종으로 취급되어 왔으며 고전적인 영지속의 분류에 문제점이 많이 있음을 시사해 주고 있다. 따라서 영지속의 분류가 진화경로에 바탕을 둔 자연분류가 되기 위해서는 형태분류 뿐만 아니라 배양 분류와 분자생물학적이 sqnstjr등 다양한 기준에 의해서 재고되어야 할 것으로 판단된다.

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Genetic Study of Soybean Sudden Death Syndrome Pathogen(Fusarium solani f. sp. glycines) isolated from Geographically Different Fields based on RFLPs of Mitochondrial DNA

  • Cho, Joon-Hyeong;J. C. Rupe
    • 한국작물학회지
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    • 제45권2호
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    • pp.143-149
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    • 2000
  • From the soils of soybean fields in Cotton Branch Station (CBS) and Pine Tree Station (PTS), Arkansas, USA, various single spore isloates of sudden death syndrome (SDS) pathogen were obtained on modified Nash & Snyder's medium (MNSM) with dilution plating technique and transferred to potato dextrose agar (PDA) medium to identify the cultural colony shape. The colony shapes of these isolates resembled F. solani isolate 171 which was white and chalky shaped on MNSM and most of them had unique form of morphology which produced white margin and blue center colony on PDA. Although, some of these isolates had more dark blue or showed slightly different color, all isolates that were selected randomly for green-house inoculation assay produced typical foliar symptoms on leaves of soybean, Hartz 6686. To determine the genetic differences among the isolates, mitochondrial DNA restriction fragment length polymorphism (RFLP) was conducted with fourty isolates from both fields, using mtDNA probes, 2U18 and 4U40, derived from Colletotrichum orbiculare. We obtained distinctive RFLPs in each treatment of restriction enzyme, EcoRI and HaeⅢ. Isolates, 11-2-5 and 14-3-1-1, from CBS and isolates, 104-3-1-2 and 701-1-5-1, from PTS showed different band patterns from 171 in both or in either treatment of restriction enzymes. Even if some of these isolates showed heterogeneous, they were more closer to 171 than PN603. And, also, rest of the thirty-six isolates had exactly same polymorphisms as 171 in each treatment of restriction enzyme. Although, some of the isolates showed the different morphological shape on PDA and slightly different band patterns on RFLPs, all of the isolates selected on MNSM due to their distinctive colony shape from other fungi produced the typical foliar symptoms on soybean leaves in greenhouse inoculation assay. It might be suggested that these isolates were not genetically different from check isolate 171 and they were unique strain of F. solani.

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Endosymbionts of Acanthamoeba Isolated from Domestic Tap Water in Korea

  • Choi, Seon-Hee;Cho, Min-Kyoung;Ahn, Soon-Cheol;Lee, Ji-Eun;Lee, Jong-Soo;Kim, Dong-Hee;Xuan, Ying-Hua;Hong, Yeon-Chul;Kong, Hyun-Hee;Chung, Dong-Il;Yu, Hak-Sun
    • Parasites, Hosts and Diseases
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    • 제47권4호
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    • pp.337-344
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    • 2009
  • In a previous study, we reported our discovery of Acanthamoeba contamination in domestic tap water; in that study, we determined that some Acanthamoeba strains harbor endosymbiotic bacteria, via our molecular characterization by mitochondrial DNA restriction fragment length polymorphism (Mt DNA RFLP). Five (29.4%) among 17 Acanthamoeba isolates contained endosymbionts in their cytoplasm, as demonstrated via orcein staining. In order to estimate their pathogenicity, we conducted a genetic characterization of the endosymbionts in Acanthamoeba isolated from domestic tap water via 16S rDNA sequencing. The endosymbionts of Acanthamoeba sp. KA/WP3 and KA/WP4 evidenced the highest level of similarity, at 97% of the recently published 16S rDNA sequence of the bacterium, Candidatus Amoebophilus asiaticus. The endosymbionts of Acanthamoeba sp. KA/WP8 and KA/WP12 shared a 97% sequence similarity with each other, and were also highly similar to Candidatus Odyssella thessalonicensis, a member of the $\alpha$-proteobacteria. The endosymbiont of Acanthamoeba sp. KA/WP9 exhibits a high degree of similarity (85-95%) with genus Methylophilus, which is not yet known to harbor any endosymbionts. This is the first report, to the best of our knowledge, to show that Methylophilus spp. can live in the cytoplasm of Acanthamoeba.

Y-Single Nucleotide Polymorphisms Diversity in Chinese Indigenous Horse

  • Han, Haoyuan;Zhang, Qin;Gao, Kexin;Yue, Xiangpeng;Zhang, Tao;Dang, Ruihua;Lan, Xianyong;Chen, Hong;Lei, Chuzhao
    • Asian-Australasian Journal of Animal Sciences
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    • 제28권8호
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    • pp.1066-1074
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    • 2015
  • In contrast to high genetic diversity of mitochondrial DNA (mtDNA), equine Y chromosome shows extremely low variability, implying limited patrilines in the domesticated horse. In this study, we applied direct sequencing and restriction fragment length polymorphism (RFLP) methods to investigate the polymorphisms of 33 Y chromosome specific loci in 304 Chinese indigenous horses from 13 breeds. Consequently, two Y-single nucleotide polymorphisms (SNPs) (Y-45701/997 and Y-50869) and one Y-indel (Y-45288) were identified. Of those, the Y-50869 (T>A) revealed the highest variation frequency (24.67%), whereas it was only 3.29% and 1.97% in Y-45288 (T/-) and Y-45701/997 (G>T) locus, respectively. These three mutations accounted for 27.96% of the total samples and identified five Y-SNP haplotypes, demonstrating genetic diversity of Y chromosome in Chinese horses. In addition, all the five YSNP haplotypes were shared by different breeds. Among 13 horse breeds analyzed, Balikun horse displayed the highest nucleotide diversity (${\pi}=5.6{\times}10^{-4}$) and haplotype diversity (h = 0.527), while Ningqiang horse showed the lowest nucleotide diversity (${\pi}=0.00000$) and haplotype diversity (h = 0.000). The results also revealed that Chinese horses had a different polymorphic pattern of Y chromosome from European and American horses. In conclusion, Chinese horses revealed genetic diversity of Y chromosome, however more efforts should be made to better understand the domestication and paternal origin of Chinese indigenous horses.

Molecular Identification of Anisakis Larvae Extracted by Gastrointestinal Endoscopy from Health Check-up Patients in Korea

  • Song, Hyemi;Jung, Bong-Kwang;Cho, Jaeeun;Chang, Taehee;Huh, Sun;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • 제57권2호
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    • pp.207-211
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    • 2019
  • Anisakiasis is a zoonotic disease induced by anisakid nematodes, and endoscopic inspection is used for a diagnosis or remedy for it. Anisakis simplex, Anisakis physeteris, and Pseudoterranova decipiens had been reported to be the major species causing human infections, particularly, in Japan. However, in Korea, recent studies strongly suggested that Anisakis pegreffii is the major species of human infections. To support this suggestion, we collected anisakid larvae (n=20) from 20 human patients who were undergone gastrointestinal endoscopy at a health check-up center in Korea, and molecular identification was performed on the larvae using PCR-RFLP analysis and gene sequencing of rDNA ITS regions and mtDNA cox2. In addition, anisakid larvae (n=53) collected from the sea eel (Astroconger myriaster) were also examined for comparison with those extracted from humans. The results showed that all human samples (100%) were identified as A. pegreffii, whereas 90.7% of the samples from the sea eel were A. pegreffii with the remaining 9.3% being Hysterothylacium aduncum. Our study confirmed that A. pegreffii is the predominant species causing human anisakiasis in Korea, and this seems to be due to the predominance of this larval type in the fish (sea eels) popularly consumed by the Korean people. The possibility of human infection with H. aduncum in Korea is also suggested.

송이 자생군락 토양 내 난배양성 세균군집의 계통학적 특성 (Phylogenetic Characteristics of viable but Nonculturable Bacterial Populations in a Pine Mushroom (Tricholoma matsutake) Forest Soil)

  • 김윤지;황경숙
    • 미생물학회지
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    • 제43권3호
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    • pp.201-209
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    • 2007
  • 송이 자생군락 토양 내 세균군집의 정량적 평가를 수행한 결과 CFDA 형광염색법을 이용해 직접 계수된 생균수는 $7.4{\pm}1.19{\times}10^8{\sim}1.07{\pm}0.17{\times}10^9cells/g$ soil로 육즙영양배지(nutrient broth, NB)에서 배양된 생균수는 CFDA 계수치의 $5{\sim}8%$로 계수되었으며, $10^{-2}$으로 희석한 NB(DNB)배지에서는 $40{\sim}47%$의 계수치를 나타내었다. 이상의 결과로부터 송이 자생군락 토양내에는 배양이 곤란한 난배양성(viable but non-culturable; VBNC)세균이 다수 존재해 있는 것으로 추정되었다. 송이 자생군락 토양내 세균군집의 계통학적 특성을 검토하기 위해 토양으로부터 직접 DNA를 추출하고 16S rDNA-ARDRA cluster 분석을 통하여 대표 clone의 16S rDNA 염기서열 분석을 수행하였다. 송이 자생군락 토양으로부터 구축된 총 115 clone은 31 ARDRA cluster로 분류되었으며, ${\alpha}-,\;{\beta}-,\;{\gamma}-$ Proteobacteria, Acidobacteria, Actinobacteria 그리고 Firmicutes의 6개 계통군이 확인되었다. 이들 계통군 중 약 85%가 Acidobacteria 계통군에 속하여 압도적인 우점군임이 확인되어 매우 독특한 계통학적 특성을 나타내었다.