• 제목/요약/키워드: RFLP.

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감자 주산지 식물기생선충 분포와 뿌리혹선층의 PCR 동정 (Occurrence of Plant-parasitic Nematodes in Major Potato Production Areas and PCR Identification of Root-knot Nematodes)

  • 조명래;이영규;김점순;유동림
    • 한국응용곤충학회지
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    • 제45권1호
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    • pp.79-85
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    • 2006
  • 국내 주요감자재배지의 식물기생선충 발생상을 밝히기 위해 2004년과 2005년에 제주, 예산, 김천, 고령, 홍천, 평창, 김제, 밀양, 남원, 강릉, 인제 등 11개 감자 주산지를 대상으로 선충속별 종류와 밀도를 조사하였다. 채집한 토양시료 50개 중 30개에서 뿌리혹선충 유충이 검출되었으며 밀도는 12-69마리/토양 100cc 정도로 비교적 낮았다. 식물기생선충류 중 Pratylenchus sp., Helicotylenchus sp, Ditylenchus sp., Tylenchus sp., and Tylenchorhynchus sp.가 지역별로 검출되었으나 대부분 밀도는 매우 낮게 나타났다. 채집한 토양을 토마토 뿌리에 접종 후 뿌리혹선충을 배양하여 PCR-RLFP에 의한 정밀동정 결과 제주, 밀양, 고령에서 채집한 뿌리혹선충 암컷에서 500 bp의 PCR 산물을 얻었으며 제한 효소 Dra I을 처리한 결과 290 및 230 bp의 밴드가 형성되어 당근뿌리혹선충(Meloidogyne hapla)으로 동정하였다.

Duchenne Muscular Dystrophy에 관한 세포유전학 및 분자유전학적 연구 (Cytogenetic and Molecular Genetic Studies on Duchenne Muscular Dystrophy)

  • 홍해숙
    • Journal of Korean Biological Nursing Science
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    • 제7권1호
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    • pp.29-46
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    • 2005
  • Purpose ; 본 연구는 X-염색체와 관련된 장애 중에서 가장 흔하고 심한 Duchenne Muscular Dystrophy(DMD)의 세포유전학 및 분자유전학적 특성을 설명하기 위해서 DMD에 영향을 받고 있는 두 가계의 13명을 대상으로 가계도 분석과 염색체 분석 및 DNA 분석을 하였다. Method ; DNA분석은 DNA probe을 이용한 Southern blotting method로써 RFLPs와 DMD유전자 부위의 exon소실 유무를 조사하여 아래와 같은 결과를 얻었다. Conclusion ; A 염색체 분석 : 말초혈액과 양수를 표본으로 High-Resolution GTG염색에서 A가계와 B가계의 염색체 분석에서 12명의 염색체는 정상 X-염색체였으나 B가계의 I-2(DMD여성)에서 46, x,-x,+t(2:x)(q 21.1 : p21.2)로 나타난다. B. DNA분석3 : 1) RFLPs의 분석 J66,XJ-1.1,754-11로써 B가계의 RELPs(Restriction Fragment Length Polymorphisms)에서 J66/Pst I은 1.7hb(E), 1.6kb(e)을 보여 주었고 XJ-1.1/Taq I은 3.6kb(F), 3.0kb(f), 754-11/EoR I은 4.2kb(G), 2.0kb(g)의 대립인자를 나타내었다. 이상의 결과를 바탕으로 영향을 받고 있는 남자 (II-2)의 haplotype는 보인자인 어머니의 한쪽 인자를 받았으며 어머니와 딸은 보인자이고 임산부의 태아는 남아였고 태아의 인자들은 그의 할아버지로부터 물려받아 DMD에 영향을 받지 않은 것으로 진단되었다. 2) DMD 유전자의 exon 소실에 대한 분석 cDNA probe 8과 cDNA probe 2b-3으로써 소실에 대한 진단은 영향을 받은 남자(II-2)는 cDNA probe 8에서 12, 7.3, 6.6, 4.2kb에 소실이 있고 cDNA 2b-3은 1.7kb에 소실에 나타났다.

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Thr-6Pro missense mutation in human lysosomal acid lipase (LAL) gene in patients with familial hypercholesterolemia in Korea

  • Hwang, Hye-Suk;Hwang, Jung-Hee;Kim, Hyun-Sup;Kim, Nam-Keun;Kim, Se-Jae;Lee, Chung-Choo;Chung, Ki-Wha
    • Journal of Genetic Medicine
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    • 제2권2호
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    • pp.65-70
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    • 1998
  • Lysosomal acid lipase (LAL) plays a central role in the intracellular degradation of neutral lipids derived from plasma lipoproteins. In this study, we investigated the missense mutation within exon 2 of human LAL gene changing of codon -6 of prepeptide from threonine to proline. The Thr-6Pro mutation was detected by the HaeIII restriction fragment length polymorphism (RFLP) and single-strand conformation polymorphism (SSCP). We analyzed the mutation in subjects with 221 unrelated randomly selected control samples and 86 patients with familial hypercholesterolemia (FH) in Korea. We observed that mutation is present with high frequency in Korea compared to other populations studied previously. The frequency of PP homozygote in the FH group was observed considerably higher than that of control. However, there was no significant difference of genotype frequency between two groups. These results, together with the fact that plasma lipids and lipoproteins levels between genotypes showed no statistical difference, suggest that the Thr-6Pro mutation in the LAL gene may have no association with the increased risk of FH development.

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Isolation and Genotyping of Toxoplasma gondii in Brazilian Dogs

  • da Silva, Jamille Rodrigues;Maciel, Bianca Mendes;de Santana Souza Santos, Luana Karla Nogueira;Carvalho, Fabio Santos;de Santana Rocha, Daniele;Lopes, Carlos Wilson Gomes;Albuquerque, George Rego
    • Parasites, Hosts and Diseases
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    • 제55권3호
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    • pp.239-246
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    • 2017
  • Strains of Toxoplasma gondii in Brazil are highly genetically diverse compared to strains from North America and Europe. Dogs are epidemiologically important because they act as sentinels for T. gondii infections in humans and are good indicators of environmental contamination. The aim of this study was to isolate and genetically characterize T. gondii strains from tissues of naturally infected Brazilian dogs. For this study, 21 blood samples were collected from dogs at the Zoonosis Control Centers of $Ilh{\acute{e}}us$ and Itabuna cities, Bahia, Brazil. The sera were examined for T. gondii antibodies using the indirect hemagglutination test. Brains and hearts of seropositive dogs were bioassayed in mice to isolate and characterize T. gondii parasites by PCR-RFLP using 10 genetic markers (SAG1, newSAG2, SAG3, BTUB, c22-8, c29-2, GRA6, PK1, APICO, and L358). However, T. gondii was isolated from only 4 (57.1%) dogs, designated TgDgBr6, 13, 17, and 21. All strains were virulent, causing clinical changes (rough hair coat, lethargy, and abdominal distention) and the death of all mice within 8-20 days after inoculation. Genetic analysis of these 4 T. gondii isolates revealed 4 distinct genotypes with different clonal lineage combinations (types I, II, and III) and 2 atypical alleles. Using PCR-RFLP with several markers, this study contributes to evaluations of the genetic diversity of strains circulating in Brazil.

Molecular Characterization of Burkholderia cepacia Complex Isolates Causing Bacterial Fruit Rot of Apricot

  • Li, Bin;Fang, Yuan;Zhang, Guoqing;Yu, Rongrong;Lou, Miaomiao;Xie, Guanlin;Wang, Yanli;Sun, Guochang
    • The Plant Pathology Journal
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    • 제26권3호
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    • pp.223-230
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    • 2010
  • The Burkholderia cepacia complex isolates causing bacterial fruit rot of apricot were characterized by speciesspecific PCR tests, recA-HaeIII restriction fragment length polymorphism (RFLP) assays, rep-PCR genomic fingerprinting, recA gene sequencing, and multilocus sequence typing (MLST) analysis. Results indicated that the isolates Bca 0901 and Bca 0902 gave positive amplifications with primers specific for B. vietnamiensis while the two bacterial isolates showed different recA-RFLP and rep-PCR profiles from those of B. vietnamiensis strains. In addition, the two bacterial isolates had a higher proteolytic activity compared with that of the non-pathogenic B. vietnamiensis strains while no cblA and esmR marker genes were detected for the two bacterial isolates and B. vietnamiensis strains. The two bacterial isolates were identified as Burkholderia seminalis based on recA gene sequence analysis and MLST analysis. Overall, this is the first characterization of B. seminalis that cause bacterial fruit rot of apricot.

Polymorphism of Insulin-like Growth Factor-I Gene in 13 Pig Breeds and its Relationship with Pig Growth and Carcass Traits

  • Wang, Wenjun;Huang, Lusheng;Chen, Kefei;Gao, Jun;Ren, Jun;Ai, Huashui;Lin, Wanhua
    • Asian-Australasian Journal of Animal Sciences
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    • 제15권10호
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    • pp.1391-1394
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    • 2002
  • The polymorphism of insulin-like growth factor-I (IGF-I) in 13 pig breeds (total n=559) was detected by PCR-Hha I- RFLP, and allele A (151 bp and 28 bp) or allele B (116 bp, 35 bp and 28 bp) were observed. In these pig breeds, it was found that European pig breeds carried high frequencies of allele B, while Chinese native pig breeds carried high frequencies of allele A. Meanwhile the role of porcine IGF-I was investigated in 117 Nanchang White pigs and 360 Large Yorkshire pigs. Eight traits about growth and carcass were recorded for analyzing the associations between IGF-I gene polymorphism and performance quantitative traits. In the Nanchang White pigs, those with AA genotype generally had higher birth weight than those with AB genotype (p<0.05), but all these genotypes had no significant effect on the other traits which had been analyzed. In Large Yorkshire pigs, those with BB genotype had higher 2 months and 6 months body weight than those with AA genotype (p<0.05), and had a thicker hind-back-fat thickness and mid-back-thickness than those with AB and BB genotypes (p<0.05). And those with BB genotype were the thinnest in Large Yorkshire. Furthermore, pigs with AA genotype had a lower lean percentage than those with AB and BB genotypes (p<0.01), and the lean percentage of those with BB genotype was the highest. Based on these results, it is possible to make the IGF-I gene locus into the application of marker-assisted selection programmes.

DdeI Polymorphism in Coding Region of Goat POU1F1 Gene and Its Association with Production Traits

  • Lan, X.Y.;Pan, C.Y.;Chen, H.;Lei, C.Z.;Hua, L.S.;Yang, X.B.;Qiu, G.Y.;Zhang, R.F.;Lun, Y.Z.
    • Asian-Australasian Journal of Animal Sciences
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    • 제20권9호
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    • pp.1342-1348
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    • 2007
  • POU1F1 is a positive regulator for GH, PRL and TSH${\beta}$ and its mutations associate with production traits in ruminant animals. We described a DdeI PCR-RFLP method for detecting a silent allele in the goat POU1F1 gene: TCT (241Ser)>TCG (241Ser). Frequencies of $D_1$ allele varied from 0.600 to 1.000 in Chinese 801 goats. Significant associations of DdeI polymorphism with production traits were found in milk yield (*p<0.05), litter size (*p<0.05) and one-year-old weight (*p<0.05) between different genotypes. Individuals with genotype $D_1D_1$ had a superior performances when compared to those with genotype $D_1D_2$ (*p<0.05). Hence, the POU1F1 gene was suggested to the potential candidate gene for superior milk performance, reproduction trait and weight trait. Genotype $D_1D_1$, characterized by a DdeI PCR-RFLP detection, was recommended to geneticists and breeders as a molecular marker for better performance in the goat industry.

Genetic Variation in MicroRNAs and Risk of Oral Squamous Cell Carcinoma in South Indian Population

  • Sushma, PS;Jamil, Kaiser;Kumar, P Uday;Satyanarayana, U;Ramakrishna, M;Triveni, B
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권17호
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    • pp.7589-7594
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    • 2015
  • Background: MicroRNAs (miRNAs) are small non-coding RNA molecules, implicated in several activities like initiation, progression and prognosis of various cancers. Single nucleotide polymorphisms (SNPs) in miRNA genes can lead to alteration in mRNA expression, resulting in diverse functional consequences. The aim of our study was to investigate the association of miR-149C>T and miR-196a2C>T SNPs with susceptibility to development of oral squamous cell carcinoma (OSCC) in South Indian subjects. Materials and Methods: 100 OSCC patients and 102 healthy controls from the general population were recruited for the study. Genetic analysis was performed by polymerase chain reaction/restriction fragment length polymorphism (PCR/RFLP) as per a standard protocol. Results: The genotype frequencies in miR-196a2 polymorphism, of TT, CT and CC in the OSCC patients were 69%,10% and 22% respectively while for control group it was 80%, 15% and 5% respectively. The CC genotype of miR196a2 polymorphism was significantly associated with oral squamous cell carcinoma. The genotype frequencies in miR-149 polymorphisms of CC, CT and TT in the oral squamous cell carcinoma (OSCC) patients were 72%, 22% and 6% respectively and for control group 88%, 12% and 0% respectively. CT and TT genotypes of miR149 polymorphism were found to be significantly associated with OSCC (p = 0.05 and 0.07). Conclusions: Our study suggests that miR-196a2C>T and miR-149C>T polymorphisms may play crucial roles in the development of OSCC in South Indian subjects.

Polymorphism of the DNA Repair Gene XRCC1 (Arg194Trp) and its role in Colorectal Cancer in Kashmiri Population: a Case Control Study

  • Nissar, Saniya;Sameer, Aga Syed;Rasool, Roohi;Chowdri, Nissar A;Rashid, Fouzia
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권15호
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    • pp.6385-6390
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    • 2015
  • Background: Genetic polymorphisms in DNA repair genes may influence individual variation in DNA repair capacity, which may be associated with risk of developing cancer. For colorectal cancer the importance of mutations in mismatch repair genes has been extensively documented. Materials and Methods: In this study we focused on the Arg194Trp polymorphism of the DNA repair gene XRCC1, involved in base excision repair (BER) and its role in colorectal cancer in Kashmiri population. A case-control study was conducted including 100 cases of colorectal cancer, and 100 hospital-based age- and sex-matched healthy controls to examine the role of XRCC1 genetic polymorphisms in the context of colorectal cancer risk for the Kashmiri population. Results: Genotype analysis of XRCC1 Arg194Trp was conducted with a restriction fragment length polymorphism (RFLP) method. The overall association between the XRCC1 polymorphism and the CRC cases was found to be significant (p < 0.05) with both the heterozygous genotype (Arg/Trp) as well as homozygous variant genotype (Trp/Trp) being moderately associated with the elevated risk for CRC [OR=2.01 (95% CI=1.03-3.94) and OR=5.2(95% CI=1.42-19.5)] respectively. Conclusions: Our results suggest an increased risk for CRC in individuals with XRCC1 Arg194Trp polymorphism suggesting BER repair pathway modulates the risk of developing colorectal cancer in the Kashmiri population.

MUTYH Association with Esophageal Adenocarcinoma in a Han Chinese Population

  • Kong, Feng;Han, Xue-Ying;Luan, Yun;Qi, Tong-Gang;Sun, Chao;Wang, Jue;Hou, Hua-Ying;Jiang, Yu-Hua;Zhao, Jing-Jie;Cheng, Guang-Hui
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권11호
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    • pp.6411-6413
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    • 2013
  • Adenocarcinoma of esophagus (AE) is a complex disease, affected by a variety of genetic and environmental factors. Much evidence has shown that the MutY glycosylase homologue (MUTYH) plays a key role in the pathogenesis of many cancers. However, there have been no reports on influence on AE in the Han Chinese population. The objective of this study was to investigate this issue. A gene-based association study was conducted using three single nucleotide polymorphisms(SNPs) reported in previous studies. The three SNPs (rs3219463, rs3219472, rs3219489) were genotyped in 207 unrelated AE patients and 249 healthy controls in a case-control study using polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). The results revealed that the genotype distribution of rs3219472 differed between the case and control groups (OR=1.66,95%CI=1.11-2.48, P=0.012), indicating that an association may exist between MUTYH and AE. These findings support a signifcant role for MUTYH in AE pathogenesis in the Han Chinese population.