• 제목/요약/키워드: DD-PCR

검색결과 83건 처리시간 0.019초

Identification of Differentially Expressed Genes Related to Intramuscular Fat Development in the Early and Late Fattening Stages of Hanwoo Steers

  • Lee, Seung-Hwan;Park, Eung-Woo;Cho, Yong-Min;Kim, Sung-Kon;Lee, Jun-Heon;Jeon, Jin-Tae;Lee, Chang-Soo;Im, Seok-Ki;Oh, Sung-Jong;Thompson, J.M.;Yoon, Du-Hak
    • BMB Reports
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    • 제40권5호
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    • pp.757-764
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    • 2007
  • Marbling of cattle meat is dependent on the coordinated expression of multiple genes. Cattle dramatically increase their intramuscular fat content in the longissimus dorsi muscle between 12 and 27 months of age. We used the annealing control primer (ACP)-differential display RT-PCR method to identify differentially expressed genes (DEGs) that may participate in the development of intramuscular fat between early (12 months old) and late fattening stages (27 months old). Using 20 arbitrary ACP primers, we identified and sequenced 14 DEGs. BLAST searches revealed that expression of the MDH, PI4-K, ferritin, ICER, NID-2, WDNMI, telethonin, filamin, and desmin (DES) genes increased while that of GAPD, COP VII, ACTA1, CamK II, and nebulin decreased during the late fattening stage. The results of functional categorization using the Gene Ontology database for 14 known genes indicated that MDH, GAPD, and COP VII are involved in metabolic pathways such as glycolysis and the TCA cycle, whereas telethonin, filamin, nebulin, desmin, and ACTA1 contribute to the muscle contractile apparatus, and PI4-K, CamK II, and ICER have roles in signal transduction pathways regulated by growth factor or hormones. The final three genes, NID-2, WDNMI, and ferritin, are involved in iron transport and extracellular protein inhibition. The expression patterns were confirmed for seven genes (MDH, PI4-K, ferritin, ICER, nebulin, WDNMI, and telethonin) using real-time PCR. We found that the novel transcription repressor ICER gene was highly expressed in the late fattening stage and during bovine preadipocyte differentiation. This information may be helpful in selecting candidate genes that participate in intramuscular fat development in cattle.

Swine Leukocyte Antigen-DQA Gene Variation and Its Association with Piglet Diarrhea in Large White, Landrace and Duroc

  • Yang, Q.L.;Kong, J.J.;Wang, D.W.;Zhao, S.G.;Gun, S.B.
    • Asian-Australasian Journal of Animal Sciences
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    • 제26권8호
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    • pp.1065-1071
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    • 2013
  • The swine leukocyte antigen class II molecules are possibly associated with the induction of protective immunity. The study described here was to investigate the relationship between polymorphisms in exon 2 of the swine DQA gene and piglet diarrhea. This study was carried out on 425 suckling piglets from three purebred pig strains (Large White, Landrace and Duroc). The genetic diversity of exon 2 in swine DQA was detected by PCR-SSCP and sequencing analysis, eight unique SSCP patterns (AB, BB, BC, CC, CD, BD, BE and DD) representing five specific allele (A to E) sequences were detected. Sequence analysis revealed 21 nucleotide variable sites and resulting in 12 amino acid substitutions in the populations. A moderate level polymorphism and significant deviations from Hardy-Weinberg equilibrium of the genotypes distribution were observed in the populations (p<0.01). The association analysis indicated that there was a statistically significant difference in the score of piglet diarrhea between different genotypes, individuals with genotype CC showed a lower diarrhea score than genotypes AB ($0.98{\pm}0.09$), BB ($0.85{\pm}0.77$) and BC ($1.25{\pm}0.23$) (p<0.05), and significantly low than genotype BE ($1.19{\pm}0.19$) (p<0.01), CC genotype may be a most resistance genotype for piglet diarrhea.

PC12 세포와 A123.7 세포에서 차별적으로 발현되는 유전자의 검색 (Screening of Differentially Expressed Genes between PC12 Cells and A123.7 Cells)

  • 백승연;양병환;채영규
    • 생물정신의학
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    • 제6권1호
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    • pp.67-73
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    • 1999
  • The cAMP-dependent protein kinase(PKA) is an intracellular enzyme with serine-threonine kinase activity that plays a key role in cell growth, differentiation, and apoptosis in eukaryotes. In order to understand the PKA signal transduction pathway regulating cell life cycle and identify its role, we focused on the characterization of up-/down-regulated genes by PKA using the differential display polymerase chain reaction. Seven differentially expressed sequence tags(DEST) have been obtained. Among these DESTs, 2 DESTs were homologous to the sequence of genes from BLAST search result. KC1-5 DEST that was up-regulated in A123.7 cells was highly corresponded to mouse apoptosis-related gene(MA-3) or mouse mRNA for topoisomerase inhibitor suppressed(TIS). MA-3 was induced in various types of apoptosis, specially in NGF-deprived apoptotic PC12 cells. TIS was down-regulated in the RVC lymphoma cells incubated with topoisomerase inhibitor that induces DNA strand breakages. PG1-1 DEST that was highly expressed in PC12 cells was corresponded to transposon Tn10 3'-end. Tnansposon Tn10 was up-regulated in differentiated myeloblastic ML-1 cells by 12-O-tetradecanoylphorbol-13-acetate. This study illuminates that MA-3/TIS was down-regulated by PKA activity, and transposon Tn10 was up-regulated by it.

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Genotype distribution and gene frequency of angiotensin I-converting enzyme in Korean population

  • Yang, Young-Mok;Park, Jong-Hwan;Moon, Eon-Soo
    • Journal of Genetic Medicine
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    • 제1권1호
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    • pp.17-22
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    • 1997
  • The angiotensin converting enzyme (ACE) is a key component of the renin-angiotensin system thought to be important in the pathogenesis of hypertension and cardiovascular diseases. Deletion polymorphism in the ACE gene may be a risk factor for myocardial infarction. The insertion/deletion (I/D) polymorphism of the ACE detected by PCR analysis appears to be associated with hypertension in Koreans and its nucleotide was subcloned into T-vector and its nucleotide sequences were determined. We also examined an association between hypertension and genetic variance of ACE. We identified the angiotensin I-converting enzyme genotype in 127 hypertensive and 189 normotensive Korean subjects. The distribution of ACE genotype II, ID, DD were 39.2%, 40.2%, 20.6% respectively and the frequency for ACE alleles I and D were 0.593 and 0.407, respectively in all subjects. The frequency of D allele in Korean males is higher than that of Korean females (male; 0.438 : female; 0.267), and the frequency of I allele in Korean females is higher than that of Korean males (female; 0.733 : male; 0.562). Genotype distributions of angiotensin I-converting enzyme genes in Korean normal adult population were different from that of Caucasians (P<0.001). There were no significant differences in genotype frequency between the hypertensive control group (n=127) and the normotensive group (n=189). We observed significant differences of ACE genotype distribution between the male group and the female group in total (P=0.001) and in hypertensive Korean subjects (P=0.013).

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Identification of Glycine max Genes Expressed in Response to Soybean mosaic virus Infection

  • Jeong, Rae-Dong;Lim, Won-Seok;Kwon, Sang-Wook;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • 제21권1호
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    • pp.47-54
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    • 2005
  • Identification of host genes involved in disease progresses and/or defense responses is one of the most critical steps leading to the elucidation of disease resistance mechanisms in plants. Soybean mosaic virus (SMV) is one of the most prevalent pathogen of soybean (Glycine max). Although the soybeans are placed one of many important crops, relatively little is known about defense mechanism. In order to obtain host genes involved in SMV disease progress and host defense especially for virus resistance, two different cloning strategies (DD RT-PCR and Subtractive hybridization) were employed to identify pathogenesis- and defenserelated genes (PRs and DRs) from susceptible (Geumjeong 1) and resistant (Geumjeong 2) cultivars against SMV strain G7H. Using these approaches, we obtained 570 genes that expressed differentially during SMV infection processes. Based upon sequence analyses, differentially expressed host genes were classified into five groups, i.e. metabolism, genetic information processing, environmental information processing, cellular processes and unclassified group. A total of 11 differentially expressed genes including protein kinase, transcription factor, other potential signaling components and resistant-like gene involved in host defense response were selected to further characterize and determine expression profiles of each selected gene. Functional characterization of these genes will likely facilitate the elucidation of defense signal transduction and biological function in SMV-infected soybean plants.

Lamivudine 복용 HIV-1 감염자에게서 내성 돌연변이 검색 (Detection of Resistance Mutation to Lamivudine in HIV-1 Infected Patients)

  • 조영걸;성흥섭;이희정;김유겸;지현숙;조군제;강문원
    • 대한미생물학회지
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    • 제35권2호
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    • pp.181-190
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    • 2000
  • To investigate resistance to lamivudine (3TC), we examined the incidence of M184V in 20 HIV-1 patients treated with 3TC for $13.1{\pm}9$ months. Fourteen of 20 patients had been exposed to zidovudine (ZDV) or didanosine (ddI) prior to 3TC therapy. Nested PCR targeting to reverse transcriptase (RT) and direct sequencing were performed for peripheral blood mononuclear cells sampled serially. There were resistance mutations to ZDV in at least 9 patients at baseline, although there was no resistance mutation to 3TC. We could detect M184V in 6 (30%) out of 20 patients. The incidence of M184V increased as the duration of therapy prolongs (13% in samples <12 months; 47% in samples ${\ge}12$ months). The frequency of mutation M184V was higher in patients with previous mutation to ZDV than in patients with wild type. Resistance mutation was not detected in 7 patients. This study shows that resistance to 3TC tends to develop rapidly in patients with baseline mutations or two drugs combination therapy than in those treated simultaneously with triple drugs. This report is the first on resistance to 3TC in Korean AIDS patients.

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Relationship between angiotensin-converting enzyme gene polymorphism and muscle damage parameters after eccentric exercise

  • Kim, Jooyoung;Kim, Chang-Sun;Lee, Joohyung
    • 운동영양학회지
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    • 제17권2호
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    • pp.25-34
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    • 2013
  • This study was conducted to investigate the relationship between ACE gene polymorphism and muscle damage parameters after eccentric exercise. 80 collegiate males were instructed to take an eccentric exercise with the elbow flexor muscle through the modified preacher curl machine for 2 sets of 25 cycles (total 50 cycles). The maximal isometric strength, muscle soreness, creatine kinase (CK), and myoglobin (Mb) were measured before exercise, and 0, 24, 48, 72, and 96 hrs after exercise. The result showed that after the eccentric exercise, the maximal isometric strength significantly decreased by more than 50% (p < 0.001) and the muscle soreness, CK, and Mb significantly increased compared to those before the exercise (p < 0.001). The ACE gene polymorphism of the subjects was classified using real-time polymerase chain reaction (real-time PCR). The result showed that it consisted of 38 cases of type II (46.4%), 33 cases of type ID (43.4%), and 9 cases of type DD (10.2%). The Hardy-Weinberg equilibrium for ACE gene polymorphism was shown to have p = 0.653, which showed that each allele was evenly distributed. Although significant differences in the changes in the maximal isometric strength, muscle soreness, CK, and Mb were found according to time course (p < 0.001), no significant differences in the changes in the maximal isometric strength, muscle soreness, CK, and Mb were found according to ACE gene polymorphism. Furthermore, no significant difference in the changes in the muscle damage parameters was found according to interaction between ACE gene polymorphism and time course (p > 0.05). In conclusion, the level of the muscle damage parameters changed in the injured muscle after eccentric exercise, but these changes in the muscle damage parameters were not affected by ACE gene polymorphism. The result of this study indicates that ACE gene is not a candidate gene that explains muscle damage.

In-silico annotation of the chemical composition of Tibetan tea and its mechanism on antioxidant and lipid-lowering in mice

  • Ning Wang ;Linman Li ;Puyu Zhang;Muhammad Aamer Mehmood ;Chaohua Lan;Tian Gan ;Zaixin Li ;Zhi Zhang ;Kewei Xu ;Shan Mo ;Gang Xia ;Tao Wu ;Hui Zhu
    • Nutrition Research and Practice
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    • 제17권4호
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    • pp.682-697
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    • 2023
  • BACKGROUND/OBJECTIVES: Tibetan tea is a kind of dark tea, due to the inherent complexity of natural products, the chemical composition and beneficial effects of Tibetan tea are not fully understood. The objective of this study was to unravel the composition of Tibetan tea using knowledge-guided multilayer network (KGMN) techniques and explore its potential antioxidant and hypolipidemic mechanisms in mice. MATERIALS/METHODS: The C57BL/6J mice were continuously gavaged with Tibetan tea extract (T group), green tea extract (G group) and ddH2O (H group) for 15 days. The activity of total antioxidant capacity (T-AOC) and superoxide dismutase (SOD) in mice was detected. Transcriptome sequencing technology was used to investigate the molecular mechanisms underlying the antioxidant and lipid-lowering effects of Tibetan tea in mice. Furthermore, the expression levels of liver antioxidant and lipid metabolism related genes in various groups were detected by the real-time quantitative polymerase chain reaction (qPCR) method. RESULTS: The results showed that a total of 42 flavonoids are provisionally annotated in Tibetan tea using KGMN strategies. Tibetan tea significantly reduced body weight gain and increased T-AOC and SOD activities in mice compared with the H group. Based on the results of transcriptome and qPCR, it was confirmed that Tibetan tea could play a key role in antioxidant and lipid lowering by regulating oxidative stress and lipid metabolism related pathways such as insulin resistance, P53 signaling pathway, insulin signaling pathway, fatty acid elongation and fatty acid metabolism. CONCLUSIONS: This study was the first to use computational tools to deeply explore the composition of Tibetan tea and revealed its potential antioxidant and hypolipidemic mechanisms, and it provides new insights into the composition and bioactivity of Tibetan tea.

표고균사 갈변과 관련된 BCR (Brown Color Repressor) 유전자 분리 (BCR (Brown Color Repressor) gene isolation related to mycelial browning of Lentinus edodes)

  • 김영호;박수철;전창성;유창현;성재모;공원식
    • 한국버섯학회지
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    • 제10권3호
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    • pp.120-128
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    • 2012
  • 표고균사에서 갈변시기를 조절하고 확인할 수 있는 유전공학적 시스템을 개발하여 톱밥재배용 표고균주를 조기에 선별할 수 있도록 표고균사가 갈변되는 동안 균사상태에서 특이적으로 발현되는 유전자를 분리하기 위하여 갈변되지 않은 균사와 갈변이 완전히 이루어진 균사에서 differential display를 실시하였다. 그 결과 이들 균사로부터 특이적으로 발현되는 두 개의 1.6kb와 1.2kb의 cDNA clone을 선발하여 염기서열을 분석하였다. 이 중 1.6kb의 cDNA단편은 Dugenia polichroa로부터 분리된 microsatellites 유전자와 100%의 상동성을 나타냈다. 그러나 1.2kb의 cDNA 단편은 3'부위에 poly A tail과 5 부위에 partial open reading frame를 가지고 있어 이를 primer로 제작한 후 갈변되지 않은 균사와 갈변이 이루어진 균사에서 RT-PCR을 실시하여 본 결과 갈변이 되지 않은 백색의 균사에서 발현이 확인되었다. 1.2kb의 cDNA 단편의 5' 부위의 염기서열 분석은 110개의 아미노산으로 구성된 partial open reading frame으로 나타났다. 이 유전자를 DNASIS database에서 상동성을 비교해 본 결과 Arabidopsis thaliata에서 분리된 dTDP-glucose 4,6-dehydratases 유전자와 DNA 수준에서는 66.7%, 아미노산 수준에서는 69.2%의 높은 상동성을 나타내었다. 갈변에 관련된 특이 유전자(BCR gene)를 확인하였다. 이 유전자는 산화 stress에 대해 저항성을 나타내는 기능을 가진 것으로 알려져 있어 표고 균사가 갈변될 때 repressor로서 작용할 것으로 생각된다. 따라서 이 유전자를 BCR (Brown Color Repressor) 유전자라고 명명하였다.

터너 증후군에서 신기형의 발생에 미치는 레닌-안지오텐신계 유전자 다형성의 영향 (Impact and Prevalence of Renin-angiotensin System Gene Polymorphism of Renal Anomalies in Turner Syndrome)

  • 박지경;정영희;이정녀;정우영
    • Childhood Kidney Diseases
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    • 제7권1호
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    • pp.52-59
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    • 2003
  • 목적 : 레닌-안지오텐신계는 신장의 성장과 발달에 중요한 역할을 하는 것으로 알려져 있으며, 안지오텐신 II는 신장형성기 동안 중요한 역할을 담당한다. 저자들은 터너 증후군 환자에서 신기형의 발생빈도를 조사하고, 신기형이 동반된 군과 동반되지 않은 군으로 분류하여 양군 사이에 레닌-안지오텐신계 유전자의 분포에 차이가 있는지를 조사하였다. 방법 : 말초혈액의 임파구를 사용하여 세포분 석학적 방법으로 진단된 33명의 터너 증후군 환자를 대상으로 신요로계의 기형여부를 초음파검사와 경정맥 요로 조영 검사, DMSA scan으로 진단하고, 더불어 안지오텐신전환효소의 유전자형, 안지오텐시노겐 유전자형, 안지오텐신 수용체유전자형의 분포를 핵산증폭법을 사용해 조사하였다. 결과 : 33명의 터너 증후군 환자 중 12명에서 신기형이 동반되어 전체적으로 36.4%의 빈도를 나타내었다. 핵형에 따른 신기형의 빈도를 살펴보면 전형적인 45,X형의 경우는 18명 중 8명에서 관찰되어 44.4%의 빈도를 나타내었고, 모자이시즘과 X 염색체구조상의 이상인 경우에는 15명 중 4명에서 관찰되어 26.7%의 빈도를 나타내어, 전형적인 45,X형의 경우에서 발생빈도가 높았으나 통계적으로 유의하지는 않았다(P>0.05). ACE 유전자형의 분포는 신기형 동반군에서 DD형이 0%, ID형이 73%, 그리고 II형이 27%이었으며, 신기형이 동반되지 않은 군에서는 DD형이 11%, ID형이 56%, II형이 33%로 양군 사이에는 ACE 유전자형의 분포는 유의한 차이가 없었다. AGT M235T 유전자형의 분포는 신기형 동반군에서 MM형이 82%, MT형이 8%, 그리고 TT형이 0%이었으며, 신기형이 동반되지 않은 군에서는 MM형이 56%, MT형이 33%, TT형이 11%로 양군 사이에는 AGT M235T 유전자형의 분포는 유의한 차이가 없었다. ATR 1 유전자형의 분포는 신기형 동반군에서 AA형이 91%, AC형이 9%, 그리고 CC형이 0%이었으며, 신기형이 동반되지 않은 군에서는 AA형이 94%, AC형이 6%, CC형이 0%로 양군 사이에는 ATR 1 유전자형의 분포도 유의한 차이가 없었다. 결론 : 터너증후군 환자에서 신기형의 동반율은 36.4%였으며, 45,X형에서 발생빈도가 높았으나 통계적으로 유의하지는 않았다. 신기형이 동반된 군과 동반되지 않은 군 사이에는 레닌-안지오텐신계 유전자의 분포는 모두 유의한 차이가 관찰되지 알았다.

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