• Title/Summary/Keyword: RFLP.

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Association of functional sequence variants of the myosin heavy chain 3 gene with muscle collagen content in pigs

  • Yong-Jun Kang;Sang-Hyun Han;Sang-Geum Kim;Su-Yeon Kim;Hyeon-Ah Kim;Yoo-Kyung Kim;Ji-Hyun Yoo;Moon-Cheol Shin;Byoung-Chul Yang;Hee-Bok Park;Jun Heon Lee;In-Cheol Cho
    • Journal of Animal Science and Technology
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    • v.65 no.3
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    • pp.511-518
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    • 2023
  • This study examined the association between functional sequence variants (FSVs) of myosin heavy chain 3 (MYH3) genotypes and collagen content in a Landrace and Jeju native pig (JNP) crossbred population. Four muscles (Musculus longissimus dorsi, Musculus semimembranosus, Musculus triceps brachii, and Musculus biceps femoris) were used for the analysis of meat collagen content, and the same animals were genotyped for the FSVs of the MYH3 gene by using PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism). Three FSVs of MYH3 genotypes were identified and had genotype frequencies of 0.358, 0.551, and 0.091 for QQ, Qq, and qq, respectively. QQ animals for the FSVs of the MYH3 genotypes showed higher collagen content in their M. longissimus dorsi (p < 0.001), M. semimembranosus (p < 0.001), M. triceps brachii (p < 0.001), and M. biceps femoris (p < 0.001) than qq homozygous animals. After the validation of this result in other independent populations, the FSVs of MYH3 genotypes can be a valuable genetic marker for improving collagen content in porcine muscles and can also be applied to increase the amount of collagen for biomedical purposes.

The variation of insulin like growth factor 2 maker is associated with growth traits in Thai native (Kradon) pigs

  • Kessara Ampaporn;Rattikan Suwannasing;Pitchayanipa Phongphanich;Supanon Tunim;Monchai Duangjinda
    • Animal Bioscience
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    • v.36 no.9
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    • pp.1350-1356
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    • 2023
  • Objective: This study was conducted to investigate polymorphisms of the melanocortin-4 receptor (MC4R) and insulin like growth factor 2 (IGF2) genes and to evaluate the growth traits affected by such polymorphisms in Thai native (Kradon) pigs. Methods: Blood samples and productive data from 91 Kradon pigs were collected. DNA was extracted and quantified, the IGF2 and MC4R genes were amplified, and the polymerase chain reaction (PCR) produces were digested using the PCR-restriction fragment length polymorphism (PCR-RFLP) technique. Genotyping was performed, and the association between genotypes and growth traits on the birth and weaning weights were evaluated. Results: The IGF2 intron7 g.162G>C variations in Kradon pigs were found in three genotypes: i) GG, ii) GC, and iii) CC. The GG genotype frequency was the highest followed by the GC and CC genotypes. The frequencies of the G and C alleles were 0.703 and 0.297, respectively. The MC4R genotype was found in only one genotype (GG). The IGF2 gene pattern was not associated with birth weight traits, whereas the IGF2 gene pattern was related to the weaning weight trait in Kradon pigs. Pigs with the CC and GC genotypes had higher weaning weights than ones with the GG genotype (p<0.001). Conclusion: Thai native Kradon pigs with the CC and GC genotypes of the IGF2 gene have higher weaning weights than pigs with the GG genotype.

Analysis of Molecular Diversity in Castanopsis sieboldii with Felt Disease Caused by Septobasidium sp. (Septobasidium sp.에 의한 구실잣밤나무 고약병의 분자학적 다양성 분석)

  • Geon-Woo Lee;Sang-Tae Seo;Byeongjin Cha;Sang-Sub Han
    • Research in Plant Disease
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    • v.29 no.4
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    • pp.420-424
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    • 2023
  • In 2020, within the Dongbaekdongsan area in Jeju Island, a Septobasidium sp. associated with a felt disease in Castanopsis sieboldii (Makino) Hatus. ex T. Yamaz. & Mashiba was identified. The symptom included the presence of brown, thin, and silk-like mycelial mats attached to the tree's bark, displaying variations in size from large to small. To induce hyphal growth, the samples collected were incubated in a moist chamber, and the newly formed hyphae were subjected to genomic DNA extractions. The nucleotide sequences of the internal transcribed spacer and small subunit rDNA genes were determined, and molecular characteristics among the isolates were investigated through polymerase chain reaction-based restriction fragment length polymorphism analysis. This Septobasidium sp. exhibited distinct morphological and phylogenetic features compared to those that were previously reported in South Korea. Consequently, this strain is taxonomically classified as a provisionally novel species of Septobasidium. Furthermore, the observed felt disease exhibited a high degree of host specificity, as it was exclusively identified in C. sieboldii without occurrence in other tree species at the time of observation.

Association of apolipoprotein E polymorphisms with serum lipid profiles in obese adolescent (비만아에서 고지혈증과 Apolipoprotein E 다형성의 관계)

  • Yoon, Jung Min;Lim, Jae Woo;Cheon, Eun Jung;Ko, Kyoung Og
    • Clinical and Experimental Pediatrics
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    • v.51 no.1
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    • pp.42-46
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    • 2008
  • Purpose : Apolipoprotein E (Apo E) plays a major role in lipoprotein metabolism and lipid transport. Many investigators have described that Apo E polymorphisms is one of the most important genetic determinants for cardiovascular disease. The purpose of this study was to evaluate the association between Apo E polymorphisms and serum lipid profiles in obese adolescent. Methods : We measured the serum concentrations of glucose, apolipoprotein (Apo) A1, Apo B, total cholesterol (TC), triglyceride (TG), HDL and LDL-cholesterol after overnight fasting in obese adolescent. Apo E polymorphisms were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results : 86 obese adolescents participated in this study. The body mass index (BMI) of participants were excess of 95 percentile by age and sex. Male to female ratio was 1.7 and mean age of study group was $16.2{\pm}1.8\;years$. Mean BMI was $27.4{\pm}2.5kg/m^2$. The frequency of ${\varepsilon}2$, ${\varepsilon}3$ and ${\varepsilon}4$ allele were 8.1%, 87.2% and 4.7% respectively. Study populations were classified into the following three genotypes 1) Apo E2 group (n=13, 15.1%) carrying either the ${\varepsilon}2/{\varepsilon}2$ or ${\varepsilon}2/{\varepsilon}3$ 2) Apo E3 group (n=65, 75.6%) carrying the most frequent ${\varepsilon}3/{\varepsilon}3$ 3) Apo E4 group (n=8, 9.3%) carrying either the ${\varepsilon}3/{\varepsilon}4$ or ${\varepsilon}4/{\varepsilon}4$. No differences were found among Apo E genotypes concerning age, sex, weight, height and BMI. Apo B and LDL-cholesterol concentrations were significantly higher in the Apo E4 group (P<0.05). No association were found between Apo E genotypes and glucose, Apo A1, TC, TG and HDL. Conclusions : We confirmed that serum concentrations Apo B and LDL-cholesterol were influenced by Apo E genotypes. Apo E polymorphisms seems to influence some alteration of lipid metabolism associated with obesity in adolescent.

Linkage Map and Quantitative Trait Loci(QTL) on Pig Chromosome 6 (돼지 염색체 6번의 연관지도 및 양적형질 유전자좌위 탐색)

  • Lee, H.Y.;Choi, B.H.;Kim, T.H.;Park, E.W.;Yoon, D.H.;Lee, H.K.;Jeon, G.J.;Cheong, I.C.;Hong, K.C.
    • Journal of Animal Science and Technology
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    • v.45 no.6
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    • pp.939-948
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    • 2003
  • The objective of this study was to identify the quantitative traits loci(QTL) for economically important traits such as growth, carcass and meat quality on pig chromosome 6. A three generation resource population was constructed from cross between Korean native boars and Landrace sows. A total of 240 F$_2$ animals were produced using intercross between 10 boars and 31 sows of F$_1$ animals. Phenotypic data including body weight at 3 weeks, backfat thickness, muscle pH, shear force and crude protein level were collected from F$_2$ animals. Animals including grandparents(F$_0$), parents(F$_1$) and offspring(F$_2$) were genotyped for 29 microsatellite markers and PCR-RFLP marker on chromosome 6. The linkage analysis was performed using CRI-MAP software version 2.4(Green et al., 1990) with FIXED option to obtain the map distances. The total length of SSC6 linkage map estimated in this study was 169.3cM. The average distance between adjacent markers was 6.05cM. For mapping of QTL, we used F$_2$ QTL Analysis Servlet of QTL express, a web-based QTL mapping tool(http://qtl.cap.ed.ac.uk). Five QTLs were detected at 5% chromosome-wide level for body weight of 3 weeks of age, shear force, meat pH at 24 hours after slaughtering, backfat thickness and crude protein level on SSC6.

Association Between the Polymorphism in the Promoter Region of Porcine A-FABP Gene and Growth Traits in Duroc Pigs (돼지 Duroc 품종에서 A-FABP promoter의 다형성과 성장형질의 연관성)

  • Han, Sang-Hyeon;Jo, In-Cheol;Lee, Jong-Eon;Kim, Hyo-Seon;Lee, Jeong-Gyu;Jeon, Jin-Tae;O, Mun-Yu;Go, Mun-Seok
    • Journal of Animal Science and Technology
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    • v.48 no.1
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    • pp.21-28
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    • 2006
  • A polymorphism was found in the promoter region of porcine adipocyte fatty acid binding protein gene(A-FABP) gene which plays a key role in the binding and transportation of free fatty acid in adipocyte and deposition of intramuscular fat. Mutation was detected a substitution(T406C) using SSCP analysis and subsequently confirmed by sequencing the fragment in Duroc pigs. This T-406C mutation might change the binding activity for transcription factor nuclear factor 1(NF1). In this population, this mutation was genotyped using HinfⅠRFLP, and found three kinds of genotypes(TT, TC, and CC) showing their frequencies of 42.3, 44.3, and 13.4%, respectively. We statistically analyzed the association between the A-FABP genotypes and growth traits and found that the body weights of the pigs containing 406C/(TC or CC) were heavier for the body weight at the age of 20 weeks than those containing genotype TT(P<0.05), but not for those at the age of 0, 3, and 10 weeks. Pigs containing genotype CC had also a higher value for the average daily gain and lower values for the date for 90kg of body weight and food conversion ratio than those of 406T/- genotype. In addition, without the significant difference of back fat thickness, there was a significant association between the existence of allele CC and lean meat and eye muscle area(P<0.05). As a result of this study, we suggest that the allele T406C in the promoter region of A-FABP gene play an important role in deposition of intramuscular fat and weight in the later growth period. This polymorphism will be an useful molecular marker for breeding of Duroc pigs.

Association Analysis of MUC5AC Promoter Polymorphism with Asthma (MUC5AC 프로모터의 유전자 다형성과 천식과의 연관성)

  • Han, Seon-Sook;Sung, Ji Hyun;Lee, Mi-Eun;Lee, Seung-Joon;Lee, Sung Joon;Kim, Woo Jin
    • Tuberculosis and Respiratory Diseases
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    • v.63 no.3
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    • pp.235-241
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    • 2007
  • Background: Airway mucus hypersecretion plays an important role in the pathogenesis of asthma, and is associated with the induction of MUC5AC expression in airway secretion. The MUC5AC gene is highly polymorphic; however, there are few studies about the association between the polymorphisms of the MUC5AC gene and asthma susceptibility or asthma phenotypes. We have investigated the association of MUC5AC promoter polymorphisms with the risk of asthma and asthma phenotypes. Methods: We determined the genotypes of the MUC5AC promoter (-1274G>A) in 78 asthma patients and in 78 age, sex-matched control individuals in the Korean population. Genomic DNAs from blood were analyzed by PCR and RFLP (restriction fragment length polymorphism) determination. We examined $FEV_1$, total eosinophil count, serum IgE level, $PC_{20}$ and the presence of atopy (by a skin test) in asthma patients. Results: The mean age of the patients was $47.7{\pm}16.1$ years and 38.5% were men, and the mean $FEV_1$ was $84.4{\pm}22.3%$ of predicted in the asthma patients. The -1274G>A polymorphism of the MUC5AC promoter in asthma patients was not significantly different as compared with normal individuals (GG 57.7%, AG 34.6% and AA 7.7% in asthma patients vs. GG 56.4%, AG 38.5% and AA 5.1% in control subject, p = 0.752, Cod). Several clinical parameters in asthma patients such as $FEV_1$, total eosinophil count, serum IgE level, $PC_{20}$ and the presence of atopy, were not associated with the -1274G>A polymorphism of the MUC5AC promoter. Conclusion: The -1274G>A single nucleotide polymorphism (SNP) frequency of the MUC5AC promoter was not associated with asthma in a Korean population.

Genetic Effects of Molecular Markers Related to Carcass Traits in Hanwoo Cattle (한우 도체형질 관련 분자표지의 유전적 효과)

  • Shin, Sung-Chul;Chung, Eui-Ryong
    • Journal of Life Science
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    • v.30 no.3
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    • pp.230-238
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    • 2020
  • Carcass traits are the most economically important traits in Hanwoo (Korean cattle). Recently, the development of the field of genomics has made it possible to identify DNA markers for the genetic evaluation of carcass and meat quality traits in beef cattle. The objective of this study was to assess the genetic effects of single nucleotide polymorphism (SNP) markers related to carcass traits by field evaluations in a commercial Hanwoo population. We evaluated 15 SNP markers (TG g.371T>C, APM1 g.1454G>A, FABP4 g.2834C>G, FABP4 g.3533T>A, FABP4 g.3691G>A, SCD g.10153A>G, SCD g.10329T >C, CPE g.601T>C, EDG1 g.166A>G, NPY g.4271T>C, GPD1 g.2766C>T, PDE1B g.17122A>G, PDE1B g.17507A>C, TNNT1 g.6650C>T, and RORC g.20152A>G) related to carcass traits in Hanwoo. Genotyping of these SNP markers was performed using PCR-RFLP analysis in Hanwoo steers (n = 1,536) to evaluate their association with carcass traits. Seven SNPs, APM1 g.1454G>, FABP4 g.3691G>A, SCD g.10153A>G, CPE g.601T>C, PDE1B g.17122A>G, TNNT1 g.6650C>T, and RORC g.20152A>G, were significantly associated with carcass traits such as marbling score (MS), backfat thickness (BF), musculus longissimus dorsi area (LDA), carcass weight (CW), meat grade (MG), meat color (MC), and maturity score (MA). The results suggest that these SNPs may be used as DNA markers for the selection of Hanwoo with higher meat quality.

Association of Melanocortin 4 Receptor (MC4R) Genotypes with Fatty Acid Compositions in an Intercross Population between Duroc and Jeju Native Pigs (제주재래돼지와 듀록 참조축군에서 Melanocortin 4 Receptor (MC4R) 유전자형과 지방산 조성간의 관련성 분석)

  • Kang, Yong-Jun;Kim, Sang-Geum;Kim, Su-Yeon;Shin, Moon-Cheol;Woo, Jae-Hoon;Kim, Nam-Young;Shin, Sang-Min;Choi, Jae-Young;Yoo, Ji-Hyun;Park, Nam-Geon;Yang, Byoung-Chul;Cho, In-Cheol
    • Journal of Life Science
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    • v.30 no.1
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    • pp.58-63
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    • 2020
  • This study was conducted to examine association between melanocortin-4-receptor (MC4R) genotypes and fatty acid (FA) composition in an F2 intercross between Duroc and Jeju (South Korea) Native pigs (JNP). Fourteen FA composition traits were measured in more than 290 F2 progeny population produced between Duroc and JNP. All experimental pigs were successfully genotyped for the MC4R c.1426A>G (p.Asp298Asn) single nucleotide polymorphism (SNP) by using Taq I PCR-RFLP methods. We detected three MC4R genotypes, AA, AG, and GG with 0.299, 0.542, and 0.159 genotype frequencies, respectively. The MC4R AA genotype animals showed higher levels in palmitic acid (C16:0, p<0.05), stearic acid (C18:0, p<0.01), eicosenoic acid (C20:1n9, p<0.05), saturated fatty acid (SFA, p<0.01) than GG homozygotes, respectively. Whereas MC4R GG genotype showed higher values in linoleic acid (C18:2n6, p<0.001), linolenic acid (C18:3n3, p<0.001), linolenic acid (C18:3n6, p<0.001), arachidonic acid (C20:4n6, p<0.001) and unsaturated fatty acid (USFA, p<0.01) than AA pigs, respectively. The MC4R GG genotype was associated with increasing USFA and decreasing SFA in the F2 progeny population produced between Duroc and JNP. Our finding suggests that the MC4R polymorphisms can be used as a valuable genetic marker for Duroc and JNP breeding programs to improve meat quality and to control FA compositions.

Correlation Between Primary Tuberculous Pleurisy and NRAMP1 Genetic Polymorphism (결핵성 흉막염 환자에서 NRAMP1 유전자 다형성에 대한 연구)

  • Kim, Je-Hyeong;Kim, Byung-Gyu;Jung, Ki-Hwan;Park, Sang-Myun;Lee, Sang-Youb;Lee, Sin-Hyung;Sin, Cheol;Cho, Jae-Youn;Shim, Jae-Jeong;In, Kwang-Ho;Yoo, Se-Hwa;Kang, Kyung-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.2
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    • pp.155-165
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    • 2000
  • Background: The phagolysosomal function of alveolar macrophage against M. tuberculosis infection is influenced by Nramp1, which is encoded by the NRAMP1 gene. There are several genetic polymorphisms in NRAMP1, and these polymorphisms affect the innate host resistance through the defect in production and function of Nramp1. To investigate this relationship, the NRAMP1 genetic polymorphism in patients with primary tuberculous pleurisy was determined. Methods: Fifty-six primary tuberculous pleurisy patient, who were diagnosed by pleural biopsy, were designated to the pleurisy group and 45 healthy adults were designated to the healthy control group. Three genetic polymorphisms of NRAMP1, such as a single point mutation in intron 4(469+14G/C, INT4), a nonconservative single-base substitution at codon 543 that changes aspartic acid to asparagine(D543N) and a TGTG deletion in the 3' untranslated region(1729+55delI4, 3'UTR), were determined. Polymerase chain reaction(PCR) and polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) were used. Results: The frequencies of mutant genotypes of INT4 and 3'UTR were significantly high in pleurisy group(p=0.001, p=0.023). But the frequencies of D543N were not significantly different between the two groups(p=0.079). The odds ratios, which are a comparison with wild genotype for determining mutant genotypes, were 8. 022(95% confidence interval=2.422-26.572) for INT4 and 5.733(95% confidence interval = 1.137~28.916) for 3'UTR ; these were statistically significant But the ratio for D543N was not significant In the combined analysis of the INT4 and 3'UTR polymorphisms, the odds ratios were 6.000(95% confidence interval = 1.461~24.640) for GC/++ genotype and 14.000(95% confidence interval=1.610~121.754) for GC/+del when compared with GG/++ homozygotes ; these were statistically significant. Conclusion: Among the NRAMP1 genetic polymorphisms, a single point mutation in intron 4(469+14G/C, INT4) and a TGTG deletion in the 3' untranslated region(1729+55del4, 3'UTR) were closely related to the primary tuberculous pleurisy.

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