• Title/Summary/Keyword: mspI

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

  • Yu, Kyong-Nae;Park, Chung-Ho
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.3
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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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    • v.19 no.12
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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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    • v.15 no.9
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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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    • v.17 no.7
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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.

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

  • 최혜선;신환성;김희종;김경수;우수진
    • Korean Journal of Microbiology
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    • v.35 no.3
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    • pp.173-179
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    • 1999
  • As a result of PCR-RFLP, the isolates used in this study were classified into five groups. Isolates 1 and 3 were included in AG-5 with 97% genetic similarity. Isolates 12 and 13 were included in AG-1 wilh 100% genetic similarily. Isolates 10 and AG-2-2 showed 97% similarity Isolates 7, 8, 11. 13, and 15 were included in AG-1. When isolates of 4, 5, 7 and 8 were restricted with Hae I. there was a single 700 bp fragment matched with AG-1. A 517 bp restriction fragment of isolate 9 was matched with AG-2-1. Based on the result of southem hybridization of genomic DNAs, all isolates restricted with Msp I showed more variable restriction differences than those restricted with Hae Ill. Isolates AG-2-1 and 9 showed 200 bp restriction fragment, and isolates 3 and AG-1 showed 1 kb restriction fragments.

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

  • Cha, In-Ho;Kwon, Jong-Jin;Park, Kwang-Kyun
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.28 no.5
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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.

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

  • Kim, Gye-Woong;Yoo, Jae-Young
    • Journal of Animal Science and Technology
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    • v.51 no.4
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    • pp.283-288
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    • 2009
  • This study was carried out to compare Msp I polymorphisms in the 3rd intron of porcine gene encoding the pituitary-1 transcription factor (POU1F1) from 286 pigs (Landrace $\times$ Yorkshire $\times$ Duroc, LYD) and to determine the associations between its genotypes and carcass traits by using the PCR-RFLP technique. The frequency of the single nucleotide polymorphism (SNP) genotype DD (84.33%) was very higher than that of CC genotype (0.75%). Allelic frequencies for C and D were 0.082 and 0.918, respectively. Each population followed the Hardy-Weinberg equilibrium. Meat colours of Hunter $L^*$ values and visual colour according to two genotypes were all significantly different. However, no significant difference in crossbred (LYD) was found between CD and DD genotypes for other traits. Therefore, this suggests that POU1F1 may be a major gene or marker for carcass traits.

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

  • 이상용;박선정;최장경
    • Korean Journal Plant Pathology
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    • v.14 no.4
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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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    • v.7 no.2
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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).

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

  • 방주희;박선정;이금희;최장경;이상용
    • Research in Plant Disease
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    • v.7 no.1
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    • pp.1-7
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    • 2001
  • An isolate of Cucumber mosaic cucumovirus(CMV) was isolated from Hydrangea macrophylla for. otaksa(Sieb. et Zucc. ) Wils. showing mosaic symptoms, and designated as Hm-CMV. Hm-CMV was characterized by the tests of host range, physical properties, serological properties, RNA and coat protein compositions, and reverse transcription and polymerase chain reaction (RT-PCR) analysis. Twelve species in 4 families were used in the host range test of Hm-CMV and could be differentiated from Y-CMV used as a control CMV by the ringspot and line pattern on inoculated leaves of several tobacco plants. Thevirus produced local lesions on inoculated leaves of Chenopodium amarticolor, C. quinoa and Vigna unguiculata. The physical properties of the virus were as follows; thermal inactivation point(TIP) was 60$\^{C}$, dilution end point (DEP) was 10$\^$-3/, and longevity in vitro (LIP) was 3∼4 days. Hm-CMV was serologically identical to Y-CMV. SDS-polyaciylamide gel electrophoresis(SDS-PAGE) showed one major protein band of about 28 kDa. In RNA or dsRNA analysis, Hm-CMV consisted of four RNA or dsRNA species, but satellite RNA was not detected. In RT-PCR using CMV-common primer and CMV subgroup I-specific primer, bothe amplified expected size of about 490 bp and 200 bp DNA fragments from Hm-CMV, respectively. Restriction enzyme analysis of the 490 bp RT-PCR products using EcoR I and Msp I showed that Hm-CMV belonged to CMV subgroup I. However, Hm-CMV could be differentiated from other CMV subgroup I isolates by RNA fingerprinting by arbitrarily primed polymerase chain reaction (RAP-PCR).

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