• Title/Summary/Keyword: RFLP.

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Effect of the single nucleotide polymorphism from cytochrome P450 (CYP2A6) gene to fatty acid composition traits in Korean native pig crossed progeny (돼지 Cytochrome P450 (CYP2A6) 유전자 내의 단일염기변이 발굴 및 고기내 불포화 지방산 조성에 미치는 영향)

  • Roh, Jung-Gun;Kim, Sang-Wook;Kim, Kwan-Suk
    • Korean Journal of Agricultural Science
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    • v.38 no.4
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    • pp.689-693
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    • 2011
  • The purpose of this study was to investigate the Cytochrome P450 (CYP2A6) gene as a candidate gene for the traits related with meat fatty acid composition traits in pigs. Porcine CYP2A6 polymorphisms were detected and PCR-RFLP was performed for genotyping of Korean native pig (n=14), Landrace (n=3), Duroc (n=3), Berkshire (n=3), Yorkshire (n=8) and F2 population composed of 202 individuals from an intercross between Korean Native pig and Yorkshire. PCR primer set amplified a 612 bp fragment of CYP2A6 and digestion of the PCR products was performed with the restriction enzymes SchI. The CYP2A6 SchI polymorphism was only found in the KNP breed. The genotype frequencies of TT, TC and CC genotypes were 0.36, 0.56 and 0.08 in the KNP respectively and the other pig breeds were fixed with CC genotype (Duroc, Landrace, Berkshire and Yorkshire). Statistical association between genotypes and fatty acid composition traits were tested in the Korean native pig and Yorkshire crossed F2 pigs. The CYP2A6 SchI polymorphism was associated with only fatty acid composition C20:3n3 level (cis11,14,17-Eicosatrienoic acid, p=0.0252). The 'T' allele was associated with lower C20:3n3 level. Further study is required to validate the genotypic association and biological consequence of the CYP2A6 gene polymorphism in pigs.

Genetic Diversity and Molecular Markers in Introduced and Thai Native Apple Snails (Pomacea and Pila)

  • Thaewnon-Ngiw, Bungorn;Klinbunga, Sirawut;Phanwichien, Kantimanee;Sangduen, Nitsri;Lauhachinda, Nitaya;Menasveta, Piamsak
    • BMB Reports
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    • v.37 no.4
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    • pp.493-502
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    • 2004
  • The genetic diversity and species-diagnostic markers in the introduced apple snail, Pomacea canaliculata and in the native Thai apple snails; Pila ampullacea, P. angelica, P. pesmei, and P. polita, were investigated by restriction analysis of COI and are reported for the first time. Twenty-one composite haplotypes showing non-overlapping distributions among species were found. Genetic heterogeneity analysis indicated significant differences between species (P < 0.0001) and within P. pesmei (P < 0.0001) and P. angelica (P < 0.0004). No such heterogeneity was observed in Pomacea canaliculata (P > 0.0036 as modified by the Bonferroni procedure), P. ampullacea (P = 0.0824-1.000) and P. polita (P = 1.0000). A neighbor-joining tree based on genetic distance between pairs of composite haplotypes differentiated all species and indicated that P. angelica and P. pesmei are closely related phylogenetically. In addition, the 16S rDNA of these species was cloned and sequenced. A species-specific PCR for P. canaliculata was successfully developed with a sensitivity of detection of approximately 50 pg of the target DNA template. The amplification of genomic DNA (50 pg and 25 ng) isolated from the fertilized eggs, and juveniles (1, 7, and 15 d after hatching) of Pomacea canaliculata was also successful, and suggested that Pomacea canaliculata and Pila species can be discriminated from the early stages of development.

The Influence of ADH1B, ALDH2 Activities and Their Combination on Drinking Behaviors of Korean Young Adults (ADH1B와 ALDH2 활성 조합이 젊은 한국인의 음주 행동에 미치는 영향)

  • Park, Jae-Young;Kim, Sie-Kyeong;Lee, Sang-Ick
    • Korean Journal of Biological Psychiatry
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    • v.17 no.1
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    • pp.26-36
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    • 2010
  • Objectives : It is well-known that Korean people show distinctive drinking behaviors depending on the gene polymorphisms of alcohol metabolizing enzymes. This study examined the gene polymorphisms of ALDH2 and ADH1B and their combination on the drinking behaviors of Korean young adults. Methods : Through a follow-up survey performed for a cohort consisting of 551 university freshmen for six years, the authors attempted to identify genetic factors affecting drinking behaviors. In 2000, drinking behaviors and scores of CAGE questionnaires were assessed and ALDH2 gene polymorphism was determined with PCR-RFLP. In 2006(n= 150), AUDIT-K was assessed in addition to the above and gene polymorphism of ADH1B was determined through SNaPshot$^{TM}$ method. Results : While ALDH2*2 allele was associated with increased degree of drinking in 2000 and 2006. When both enzymes were active, the possibility to be classified into the risk group for alcohol dependence such as AUDIT-K(>12), and CAGE(>2) was high. Conclusion : The ALDH2 genotype had a significant effect on drinking behavior and degree of drinking during early adulthood. However, the combination of the active form of ADH1B and the active form of ALDH2 can be risk factor for problem drinking.

Analysis of $LH{\beta}$ Exon 3 (Gly102Ser) Gene Mutation in Infertile Patients with Endometriosis and Polycystic Ovary Syndrome (PCOS) (자궁내막증과 다낭성 난포증후군 불임환자에서 $LH{\beta}$ Exon 3 (Gly102Ser) 유전자의 돌연변이 분석)

  • Kim, Nam-Keun;Lee, Eu-Gene;Cho, Min-Soon;Nam, Yoon-Sung;Chung, Hyung-Min;Chung, Ki-Wha;Oh, Yu-Kyoung;Ko, Jung-Jae;Cha, Kwang-Yul
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.3
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    • pp.291-294
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    • 2000
  • 연구목적: 본 연구는 자궁내막증과 다낭성 난포증후군 불임환자들을 대상으로 $LH{\beta}$ exon 3 (Gly102Ser) 유전자의 돌연변이를 탐색하고자 시도하였다. 연구재료 및 방법: 그 대상으로 26명의 자궁내막증 환자와 52명의 다낭성 난포증후군 환자 그리고, 50명의 출산 경험이 있는 건강한 여성을 대조군으로 사용하였다. 이들을 대상으로 한 돌연변이 탐색은 PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) 방법으로 수행되었다. 결과 : 그 결과 자궁내막증과 다낭성 난포증후군 환자 및 출산 경험이 있는 건강한 여성에서 그 변이형이 나타나지 않았다. 결론: 따라서, 자궁내막증과 다낭성 난포증후군 불임환자의 $LH{\beta}$ exon 3 돌연변이형은 중국인 집단에만 존재할 가능성이 높으며, 더 많은 불임환자들을 대상으로 한 연구가 요구된다.

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Mitochondrial DNA Variations among Three Species of Korean Planorbid Snails : Gyraulus convexiusculus, Hippeutis cantori and Segmentina hemisphaerula (한국산 또아리물탈행이과 3종 내의 미토콘드리아 DNA 변이)

  • Chung, Pyung-Rim;Younghun Jung;Jung, Eung-Kyung;Kim, Dae-Soon
    • The Korean Journal of Malacology
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    • v.10 no.1
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    • pp.27-37
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    • 1994
  • 한국산 담수 또아리물달팽이과(Planorbidae)에 속하는 또아리물달팽이(Gyraulus convexiusculus),수정또아리물달팽이(Hippeutis cantori) 및 배꼽또아리물달팽이(Segmentina hemisphaerula)3종에 대한 종간 유전적 변이와 이들 상호간의 분류학적 유연 관계를 생화학적 측면에서 밝히고자 하였다. 즉, 모계유전으로 자손에 유전되고 있는 미토콘드리아 DNA(mitochondrial DNA; mt DNA)의 변이를 보기위하여 제한효소(restriction enzyme)를 처리하고 잘라진 mtDNA절편들을 상호 비교하는 restriction fragmint length polymorphism(RFLP)기법을 응용하였다. 본 실험에서 10개의 제한효소 중 CIa I, Dra I, Eco RI, Hin dIII, Kpn I및 pst I의 6개 제한효소에서 좋은 결과를 얻어 종간의 공통절편(shated fragmints)을 비교하였고, 염기분화율(nucleotide divergince rate)을 각각 측정하였다. 미토콘드리아 DNA 크기(genome size)는 또아리물달팽이가 12.08 kb, 수정또아리물달팽이가 14.4 kb, 그리고 배꼽또아리물달팽이가 12.93 kb로 관찰되었다. 염기분화율(p)는 또아리물달팽이/수정또아리물달팽이 군에서 p=12.7%, 배꼽또아리물달팽이와 상기 2종군 사이의 염기분화율은 P=56.6%여서 배꼽 또아리물달팽이류는 타 2종보다 그 분화율이 매우 높음을 알 수 있었다. 이상의 결과로 보아 분류군(taxa)의 mtDNA 변이에 의한 RELP기법이 앞으로 한국산 담수 패류 연구에 널리 응용될 수 있음이 확인되었다.

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QTL Analysis of Soybean Seed Weight Using RAPD and SSR Markers

  • Chung, Jong-Il;Ko, Mi-Suk;Kang, Jin-Ho
    • Plant Resources
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    • v.3 no.3
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    • pp.184-193
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    • 2000
  • Soybean [Glycine max (L.) Merr.] seed weight is a important trait in cultivar development. Objective of this study was to identify and confirm quantitative trait loci (QTLs) for seed weight variation in the F2 and F2:3 generations. QTLs for seed weight were identified in F2 and F2:3 generations using interval mapping (MapMaker/QTL) and single-factor analysis of variance (ANOVA). In the F2 plant generation (i.e., F3 seed), three markers, OPL9a, OPM7a, and OPAC12 were significantly (P<0.01) associated with seed weight QTLs. In the F2:3 plant row generation (i.e., F4 seed), five markers, OPA9a, OPG19, OPL9b, OPP11, and Sat_085 were significantly (P<0.01) associated with seed weight QTLs. Two markers, OPL9a and OPL9b were significantly (P<0.05) associated with seed weight QTLs in both generations. Two QTLs on USDA soybean linkage group C1 and R were identified in both F2 and F2:3 generations using interval mapping. The linkage group C1 QTL explained 16% of the variation in seed weight in both generations, and the linkage group R QTL explained 39% and 41% of the variation for F2 and F2:3 generation, respectively. The linkage group C2 QTL identified in F2:3 generation explained 14.9% of variation. Linkage groups C1, C2 and R had previously been identified as harbouring seed size QTLs. The consistency of QTLs across generations and populations indicates that marker-assisted selection is possible in a soybean breeding program.

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The Effect of Trp64Arg Polymorphism in the ${\beta}_3$-Adrenergic Receptor Gene on Blood Pressure, Body Composition and Bone Mineral Density in Athletes (운동선수의 베타3-아드레날린성 수용체 유전자의 Trp64Arg 다형성이 혈압, 신체조성 및 골밀도에 미치는 영향)

  • Jung In Geun;Oh Sang Duk;Kim Tae Wook;Kang Byung Yong;Ha Nam Joo;Ha Nam Joo
    • YAKHAK HOEJI
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    • v.49 no.1
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    • pp.44-50
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    • 2005
  • The purpose of this study was to investigate the relationship between Trp64Arg polymorphism in the ${\beta}_3$-adrenergic receptor gene and complex phenotypes such as blood pressure, body compositions and bone parameters in young men about 20 years, and to collect the fundamental data in designing the exercise program. Eighty healthy young men including 41 controls and 39 athletes were recruited, Trp64Arg polymorphism in the ${\beta}_3$-adrenergic receptor gene was genotyped by PCR-RFLP method. By association study, there were no significance in genotype and allele frequencies of Trp64Arg polymorphism in the ${\beta}_3$-adrenergic receptor gene between controls and athletes, respectively (p>0.05). When the relationship between physiological parameters and Trp64Arg polymorphism in the ${\beta}_3$-adrenergic receptor gene was tested, this polymorphism was significantly associated with 3th lumber and left femoral neck Z-score values in controls (p<0.05), but these associations were not detected in athletic groups (p>0.05). It is likely that Trp64Arg polymorphism in the ${\beta}_3$-adrenergic receptor gene is a genetic marker for the bone mineral density index in young men, but environmental factors such as exercise modify the significant effect of this polymorphism. Thus, our results suggest that Trp64Arg polymorphism in the ${\beta}_3$-adrenergic receptor gene may be applicable as a predictive marker for osteoporosis in Korean young men, and regular exercise may prevent the disadventageous effect of this polymorphism for bone mineral density in male athletic group.

Pharmacokinetics and Bioequivalence Evaluation of Risperidone in Healthy Male Subjects with Different CYP2D6 Genotypes

  • Cho, Hea-Young;Lee, Yong-Bok
    • Archives of Pharmacal Research
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    • v.29 no.6
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    • pp.525-533
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    • 2006
  • The aim of this study was to evaluate the bioequivalence of risperidone in healthy male subjects representing different CYP2D6 genotypes with respect to risperidone, 9-hydroxyrisperidone (9-OH-risperidone), and active moiety. A total of 506 Korean subjects were genotyped for $CYP2D6^*10$ by means of allele-specific polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Based on the genotype analysis, 24 subjects, 7 homozygous for $CYP2D6^*1$ for $^*10$, and 7 heterozygous for $^*10$, were recruited and received a single oral dose of 2 mg risperidone tablet in this study. Serum concentrations of risperidone and 9-OH-risperidone up to 48 h were simultaneously determined. There were no significant differences of the active moiety, risperidone, and 9-OH-risperidone between the two preparations in AUC_{0-{\propto}}$ and $C_{max}$. The 90% confidence intervals (Cls) for the ratio of means of the log-trans-formed AUC_{0-{\propto}}$ and $C_{max}$ for the active moiety, risperidone, and 9-OH-risperidone were all within the bioequivalence acceptance criteria of 0.80-1.25. The $CYP2D6^*10$ allele particularly was associated with higher serum concentrations of risperidone and the risperidone/9-OH-risperidone ratio compared with the $CYP2D6^*1$ allele. The results demonstrate that the two preparations of risperidone are bioequivalent and it can be assumed that they are therapeutically equivalent and exchangeable in clinical practice. Furthermore, the pharmacokinetic parameters of risperidone and the risperidone/9-OH-risperidone ratio are highly dependent on the CYP2D6 genotypes.

Phenotype Based on Pharmacokinetic/Pharmacodynamic Parameters and Genotype Correlations of Immunosupressants (면역억제제의 약물속도론적/약력학적 파라미터에 기초한 표현형과 유전형의 상관성)

  • Lee, Yong-Bok;Cho, Hea-Young
    • Journal of Pharmaceutical Investigation
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    • v.37 no.6
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    • pp.369-376
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    • 2007
  • Cyclosporine (CsA) and tacrolimus (FK506) have a narrow therapeutic range, and their pharmacokinetic (PK) characteristic varies among individual. They are also substrates for cytochrome P450 (CYP) 3A4, 3A5 genes, and P-glycoprotein, the product of the multidrug resistance 1 (MDR1). The aims were to investigate the relationship between CYP3A and MDR1 genotypes and their PK parameters among healthy subjects. We investigated the genotype for CYP3A and MDR1 gene in human using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. After oral administration of CsA and FK506 (100 mg and 1 mg, respectively), whole blood samples were taken up to 24 hours. Blood CsA and FK506 concentrations were measured by LC/MS/MS. Each PK parameters were compared using Kruskal-Wallis test according to the CYP3A and MDR1 genotype. We found that the values of AVC for CsA were significantly different among CYP3A5 and MDR1 exon 26 (C3435T) genotypes (P=0.037 and P=0.049). On the other hand, the AUC for FK506 was significantly different only among CYP3A5 genotypes (P=0.013). The results clearly demonstrate the effects of CYP3A5 and MDR1 exon 26 on Cys and FK506 disposition.

Relationship between Stress Gene Polymorphisms and Litter Size by AI in Pigs

  • Jin, H.J.;Kim, I.C.;Wee, M.S.;Yeon, S.H.;Kim, C.D.;Lee, S.S.;Cho, C.Y.;Cho, S.R.;Son, D.S.;Park, C.K.;Li, Z.D.
    • Journal of Embryo Transfer
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    • v.22 no.4
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    • pp.257-263
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    • 2007
  • This study was performed to investigate the relationship between PSS-HSP70 gene polymorphism and artificial insemination (AI) reproductivity in the pigs. The RFLP polymorphism of PSS and the SSCP polymorphisms of HSP70 K1, K3 and K4 PCR product were detected different patterns. In the experiment for AI of fresh semen, spring and fall season showed higher litter size born of 10.89 head than 10.47 head of summer season. Landrace was showed higher litter size of 9.96 head than that of Duroc and Yorkshire (p<0.05). Stress relating PSS and HSP70 polymorphism of PSS-Normal, HSP70 K1-BB, K3-AB, K4-AA showd a highest litter size born of 10.97 head and litter size born alive of 10.69 head than that of the other polymorphisms(p<0.05). In the experiment for AI of frozen semen, effects of season and pig breeds were not showed for litter size born. The stress relating polymorphism of PSS-Carrier, HSP70 K1-BB, K3-BB, K4-AB showed highest litter size born of 11.29 head and litter size born alive of 10.82 head and PSS-Normal, HSP70 K1-BB, K3-AB, K4-AA showed the lowest litter size born of 8.48 head and litter size born alive of 7.33 head than that of the other polymorphisms(p<0.05). These results suggest that AI litter size born for the stress of forzen thawed semen may be affected by PSS and HSP70 polymorphism in pigs.