• 제목/요약/키워드: MspI

검색결과 117건 처리시간 0.029초

rpoB 유전자의 PCR-RFLP를 이용한 Mycobacterium 균종 동정의 유용성 (Identification of Mycobacterium species by rpoB Gene PCR-RFLP)

  • 유경래;박정오
    • 대한임상검사과학회지
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    • 제38권3호
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    • pp.158-165
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    • 2006
  • Although Mycobacterium tuberculosis complex strains remain responsible for the majority of diseases caused by mycobacterial infections worldwide, the increase in HIV infections has allowed for the emergence of other non-tuberculous mycobacteria as clinically significant pathogens. However, Mycobacterium species has a long period of incubation, and requires serious biochemical tests such as niacin, catalase, and nitrate test that are often tedious. The development of rapid and accurate diagnostics can aid in the early diagnosis of disease caused by Mycobacterium. The current DNA amplification and hybridization methods that have been developed target several genes for the detection of mycobacterial species such as hps65, 16S rDNA, rpoB, and dnaj. These methods produce rapid and accurate results. In this study, PCR-restriction fragment length polymorphism analysis(PCR-RFLP) based on the region of the rpoB gene was used to verify the identification of non-tuburculosis Mycobacterium species. A total of 8 mycobacterial reference strains and 13 clinical isolates were digested with restriction enzymes such as Msp I in this study. The results of using this process clearly demonstrated that all 13 specimens were identified by rpoB gene PRA method. The PCR-RFLP method based on the rpoB gene is a simple, rapid, and accurate test for the identification of Mycobacterium.

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Association of Polymorphisms in the Bovine Leptin Gene with Ultrasound Measurements for Improving in Korean Cattle

  • Kong, H.S.;Oh, J.D.;Lee, S.G.;Hong, Y.S.;Song, W.I.;Lee, S.J.;Kim, H.C.;Yoo, B.H.;Lee, H.K.;Jeon, G.J.
    • Asian-Australasian Journal of Animal Sciences
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    • 제19권12호
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    • pp.1691-1695
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    • 2006
  • The identification method that inflects real time ultrasound (RUT) and the potential application of marker assisted selection (MAS) for improvement of a cow population of Hanwoo (Korean Native cattle) was studied. The averages of RUT longissimus muscle area, RUT fat thickness, and RUT marbling score scanned at the 13th rib were 55.78 $cm^2$, 3.70 mm and 3.83 scores, respectively. We investigated the effects of the two SNPs (Kpn2 I and Msp I) in the leptin gene on carcass traits for Hanwoo cows by using ultrasound measurements. Genotype CC of the Kpn2 I had a significantly higher effect on back fat thickness (4.23 mm) and longissimus muscle area (57.57 $cm^2$) than genotype TT (3.14 mm, 53.93 $cm^2$, respectively, p<0.05). Genotype AA of the Msp I had a significantly higher effect only on marbling score (5.37) than genotype AB (3.57, p<0.05) and BB (3.37, p<0.05). Significant effects of SNPs in the leptin gene were found for the ultrasound measures of body composition in live cattle.

Lack of Association Between CYP1A1 Polymorphisms and Risk of Bladder Cancer: a Meta-analysis

  • Lu, Yu;Zhang, Xiao-Lian;Xie, Li;Li, Tai-Jie;He, Yu;Peng, Qi-Liu;Deng, Yan;Wang, Jian;Qin, Xue;Li, Shan
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권9호
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    • pp.4071-4077
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    • 2014
  • Background: The effects of CYP1A1 gene polymorphisms on the risk of bladder cancer (BC) remain controversial. We carried out a meta-analysis to clarify the role of CYP1A1 gene polymorphisms in BC. Material and Methods: A comprehensive literature search was conducted up to November 20, 2013. Odds ratios (ORs) with 95% confidence intervals (CIs) were used to estimate the strength of the association. Meta-regression, subgroup analysis, sensitivity analysis and publication bias were also performed. Results: Eight studies involving 1,059 BC cases and 1,061 controls were included. The meta-analysis showed that there was no significant association between the two common mutations of CYP1A1 and BC risk. For the I1e462Val A/G polymorphism with GG vs. AA the OR was 1.47 (95 % CI= 0.70-3.07, P =0.308). For the MspI T/C polymorphism, though a slight trend was found this was not statistically nonsignificant (CC vs.TT, OR = 1.24, 95 % CI= 0.98-1.58, P =0.078). Subgroup analyses by ethnicity also found no obvious association between CYP1A1 and BC risk. Conclusion: The present meta-analysis suggests that CYP1A1 polymorphism is not associated with bladder cancer risk.

Nucleotide Sequencing and PCR-RFLP of Insulin-like Growth Factor Binding Protein-3 Gene in Riverine Buffalo (Bubalus bubalis)

  • Padma, B.;Kumar, Pushpendra;Choudhary, V.;Dhara, S.K.;Mishra, A.;Bhattacharya, T.K.;Bhushan, B.;Sharma, Arjava
    • Asian-Australasian Journal of Animal Sciences
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    • 제17권7호
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    • pp.910-913
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    • 2004
  • Insulin-like growth factor binding protein-3 (IGFBP-3) gene is a structural gene associated with the growth and development of the animals. The present investigation was carried out to unravel nucleotide sequence and polymerase chain reactionrestriction fragment polymorphism (PCR-RFLP) of IGFBP-3 gene in buffalo. Genomic DNA was isolated from a total of 157 animals belonging to Murrah, Surti, Jaffarabadi and Nagpuri breeds of Indian riverine buffalo. A 655 bp of IGFBP-3 gene was amplified in all the breeds and amplicons were digested with Hae III, Taq I and Msp I restriction enzymes. On digestion with Hae III yielded single restriction pattern of 8 fragments of sizes 201, 165, 154, 56, 36, 19, 16 and 8 bp in all the animals studied. Similarly Taq I and Msp I also revealed single restriction pattern yielding fragments of sizes 240 and 415 bp and 145 and 510 bp, respectively. This shows nonpolymorphic nature of restriction sites in buffalo. Nucleotide sequencing of 587 bp of IGFBP-3 gene in Murrah buffalo was done and submitted to the GenBank (Accession No. AY304829). Nucleotide sequencing revealed an addition of 4 bases in the intronic region as compared to cattle.

각종 작물로부터 분리한 Rhizoctonia solani 균주의 RFLP 및 PCR-RFLP를 이용한 분자계통한 특성 구명 (Molecular Systematics of Rhizoctonia solani Isolates from Various Crops with RFLP and PCR-RFLP)

  • 최혜선;신환성;김희종;김경수;우수진
    • 미생물학회지
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    • 제35권3호
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    • pp.173-179
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    • 1999
  • Rhizoctonia solani 분류를 위해 균사의 anastomosis group과 grouprks의 유전적 차이의 분석을 위해 rDNA 의 PCR-RFLP 와 RFLP를 실시하였다. rDNA 의 PCR-RFLP 결과 R. solani 는 크게 5 group 으로 나뉘어졌다. 균주 번호 1번과 3번은 AG-5 그룹과 0.976의 높은 상동성을 보였고, 12번, 13번은 AG-2-2와 0.976의 상동성을 보였다. 균주번호 7,8,11,13,15 는 AG-1에 속하였다. 제한효소 HaeIII로 절단하였을 때 균주번호 4,5,7,8은 700 bp에서 하나의 밴드가 나타나 AG-1에 속하였고, 균주번호 9는 AG-2-1과 517 bp에서 하나의 밴드가 나타났다. 도한 제한효소 Msp I을 사용하여 southern blotting을 한 결과 더 다양한 절단양상을 보이며 AG 간의 차이가 나타났다. AG-2-1과 균주번호 9는 200 bp에서 하나의 밴드가 나타났다. 균주 3,4,5,10,11,13은 AG-1과 1kb에서 하나의 밴드로 나타났다.

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한국인 구강 편평세포암에서 Glutathione S-transferase와 CYP1A1 유전자의 다형성 (GENETIC POLYMORPHISMS OF THE GLUTATHIONE S-TRANSFERASE AND CYP1A1 GENES IN KOREAN ORAL SQUAMOUS CELL CARCINOMA)

  • 차인호;권종진;박광균
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제28권5호
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    • pp.364-371
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    • 2002
  • Many chemical compopunds are converted into reactive electrophilic metabolites by the oxidative(Phase I) enzymes, which are mainly cytochrome P-450 enzyme(CYPs). Phase II conjugating enzymes, such as glutathione S-transferase(GST), usually act as inactivation of enzymes. Genetic polymorphisms have been found to be associated with increased susceptibility to cancer of the lung, bladder, breast and colorectal. Many of the polymorphic genes of carcinogen metabolism show considerably different type of cancer among different ethnic groups as well as individuals within the same group. The aim of this study is (1) to establish the frequencies of genetic polymorphisms of GSTM1 and CYP1A1 in Korean oral squamous cell carcinoma(SCC), (2) to associate oral SCC with the risk of these genetic polymorphisms. The genetic polymorphisms of the GSTM1 and the CYP1A1 genes among 50 Korean oral SCC were analyzed using polymerase chain reaction(PCR). The results suggest that the homozygote and the mutant type of CYP1A1 MspI polymorphisms may be associated with genetic susceptibility to oral SCC in Korean. A combination of the GSTM1 null type with the homozygote(m1/m1), and the mutant(m2/m2) type of CYP1A1 MspI polymorphisms showed a relatively high risk of oral SCC in Korean. In the smoking group, the GSTM1 wild genotype may be the high risk factor of oral SCC in Korean. These data coincide with the hypothesis which states that different susceptibility to cancer of genetic polymorphisms exist among different ethnic group and different types of human cancer.

돼지의 POU1F1 Intron 3영역 유전자에 따른 도체특성 분석 (Analysis of Carcass Characteristics in the 3rd Intron of Pig POU1F1 Gene)

  • 김계웅;유재영
    • Journal of Animal Science and Technology
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    • 제51권4호
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    • pp.283-288
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    • 2009
  • 본 연구는 비육돈 268두를 대상으로 13번 염색체 중 POU1F1 intron 3영역의 유전자 Single Nucleotide Polymorphism(SNP)은 Msp I 제한효소에 의해서 구분된다. 직접 제작한 Primer로 POU1F1 유전자 중 intron 3영역을 증폭할 수 있도록 제작하였고, 823bp의 증폭산물을 기대할 수 있었다. 그 SNP 유전자형을 3개 군으로 분류하여 빈도 추정과 함께 육질관련 형질과의 연관성을 분석한 결과는 다음과 같다. POU1F1 SNP의 DD 유전자형은 84.33%로 가장 높은 출현율을 보인 반면에 CC 유전자형은 0.75%로 매우 낮게 나타났다. 대립유전자 D 빈도는 0.918로 매우 높게 출현하였으나, C 빈도는 0.082로 매우 낮게 출현하였다. Hardy-Weinberg 법칙에 따른 적합도 검정에서는 유의성 없이(p>0.05) 유전적 평형을 이루고 있는 집단임을 알 수 있었다. CD와 DD 유전자형 집단에서 등지방두께, 도체중, Hunter a 및 b값, 그리고 pH에서는 모두 유의한 차이를 보이지 않았으나, Hunter $L^*$값은 CD(37.26) 보다 DD(40.15) 유전자형 개체군에서 짙게 나타났고(p<0.05), 육색도판에 의한 육색은 DD(3.91) 보다 CD(5.09) 유전자형 개체에서 짙게 나타났다((p<0.001). 따라서 이러한 POU1F1 유전자의 SNP 유전자형은 돼지의 성장이나 도체관련 형질과의 연관성을 더욱 깊이 있게 탐색하여 명확한 구명으로 우량 돈육 생산을 위한 DNA marker로 활용되어야 할 것으로 사료된다.

PCR기법을 이용한 오이 모자이크 바이러스 개나리 분리주(CMV-Fk)의 동정과 구분 (Identification and Differentiation of Cucumber Mosaic Virus Isolated from Forsythia koreana (CMV-Fk) Using PCR Techniques)

  • 이상용;박선정;최장경
    • 한국식물병리학회지
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    • 제14권4호
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    • pp.308-313
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    • 1998
  • Reverse transcription and polymerase chain reaction (RT-PCR) techiniques were used to identification and differentiation of cucumber mosaic virus isolated from Forsythia koreana (CMV-Fk). RT-PCT used by two set of 20-mer primers one was CMV-common primers and another was CMV subgroup I-specific primers designed in a conserved region of the 3' end of CMV RNA3, amplified about 490 bp and 200 bp DNA fragments from CMV-Fk, respectively. CMV could be detected by RT-PCR at a dilution as low as 10-4 in forsythia crude sap extracts. Restriction enzyme analysis of RT-PCR products using EcoRI and MspI showed that CMV-Fk belonged to CMV subgroup I. But, analysis of RNA fingerprinting by arbitrarily primed polymerase chain reaction (RAP-PCR) showed heterogeneity of RNA3 between CMV-Fk and CMV-Y as a member of subgroup I.

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Variation of Cannonical Sentence Structure in Korean & Japanese Dialects & its Implication

  • Khym, Han-gyoo
    • International Journal of Internet, Broadcasting and Communication
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    • 제7권2호
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    • pp.142-148
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    • 2015
  • The main purpose of this squib is to provide a new principled account for variation of canonical sentence structure in Korean and Japanese based on the linguistic data commonly observed in some dialects of Korean and Japanese. Unlike the English case in which Comp(lementizer) such as 'that' in an embedded clause freely drops as far as the ECP (Lasnik & Saito 1992) is obeyed, some dialects of both Korean and Japanese show interesting linguistic data very different from those of English, thereby leading us to reasonably doubt the traditionally-accepted paradigm of the canonical sentence structure of CP for all languages. In this squib I propose, based on Korean & Japanese dialects and by developing the Minimal Structure Principle (MSP) ($Bo{\check{s}}kovi{\acute{c}}$ 1997, p. 25), that the cannonical structure of a sentence is not fixed, from the beginning at all, to be one single maximal category, CP. Instead, it should be decided to be either CP or IP, based on the feature of [${\pm}$markedness] and MSP, and the marked (or non-cannonical) embedded sentence needs to satisfy ECP for adjacency (or feature-licensing by the matrix verb in the MP terminology).

수국에서 분리한 Cucumber mosaic virus의 특성 (Characterization of Cucumver mosaic virus Isolated from Hydrangea macrophylla for. otaksa (Sieb. et Zucc) Wils.)

  • 방주희;박선정;이금희;최장경;이상용
    • 식물병연구
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    • 제7권1호
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    • pp.1-7
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    • 2001
  • 1998년 수원 근교에서 채집한 전형적인 모자이크 병징을 나타내는 수국(Hydragea macrophylla for. otaksa)으로부터 CMV를 분리하고, Hm-CMV라 명명하였다. 기주실험, 물리적 실험, 혈청학적 성질, RNA와 coat protein의 성질, RT-PCR 및 RAP-PCR 분석을 통하여 Hm-CMV의 특성을 분석하였다. 12종의 CMV 지표식물에서 실시한 기주반응실험의 결과, 지금까지 보고된 CMV 계통들의 반응과 특징적인 차이는 인정되지 않았다. Hm-CMV의 물리적 성질은 내열성에서 6$0^{\circ}C$를 보여 기존 CMV들 보다 낮았다. 혈청학적으로 Hm-CMV는 Y-CMV와 융합하는 subgroup I CMV로 분석되었다. SDS-PAGE로부터에 Hm-CMV의 외피단백질은 28 kDa의 band가 확인되었으며, 4종의 게놈 RNA는 Y-CMV와 같은 분자량을 나타냈으나, 위성 RNA는 존재하지 않았다. 수국의 이병엽에서 분리한 dsRNA의 분석 결과도 Y-CMV와 같은 패턴을 보였다. Hm-CMV의 외피단백질유전자에 대한 RT-PCR 분석 결과, 예상된 분자크기의 DNA 증폭이 인정되었으며, PCR 산물을 이용한 EcoR I 및 Msp I을 처리한 결과는 subgroup I CMV의 특성을 나타냈다. 그런, RAP-PCR의 결과, Hm-CMV는 subgroup I내의 다른 계통들과 구분되었다.

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