• Title/Summary/Keyword: RFLP.

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Screening of BCL-2 associated X protein gene polymorphism associated with scrotal hernia in domesticated swine using polymerase chain reaction-restriction fragment length polymorphism

  • Manalaysay, Jessica G.;Antonio, Nathaniel D.;Apilado, Ralph Lorenz R.;Bambico, Joseph F.;Mingala, Claro N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.2
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    • pp.262-266
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    • 2017
  • Objective: This study was conducted to screen scrotal hernia in domesticated swine from selected breeders in the Philippines. This defect is associated with a cytosine to thymine mutation in the BCL-2 associated X protein (BAX) gene of swine. Methods: Genetic screening was done by DNA extraction followed by amplification and digestion using polymerase chain reaction-restriction fragment length polymorphism, amplifying the 416 bp region of the BAX gene that was subjected to digestion using the Ear I enzyme. Sequencing was also conducted to validate the results. Results: Results revealed that out of 538 samples tested, 411 (76.4%) of the samples were found to be normal whereas the remaining were carriers of the mutation in which 80 (14.9%) were heterozygous mutants and 47 (8.7%) were homozygous mutants. Pietrain breed was found to have the highest incidence. Conclusion: Having a scrotal hernia eliminates the chances of using the boar as a breeder stock because the following generations arising from it would most likely exhibit herniation. It is therefore advised to establish a genetic screening method for Scrotal Hernia in the Philippines to eliminate the negative gene from the herd.

The Relationship between Taq I A Dopamine $D_2$ Receptor Polymorphism and Therapeutic Response to Antipsychotics in Schizophrenic Patients (정신분열병환자에서 Taq I A 도파민 $D_2$ 수용체 다형성과 항정신병약물의 치료반응과의 연관)

  • Kang, Cheol Joong
    • Korean Journal of Biological Psychiatry
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    • v.12 no.2
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    • pp.107-113
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    • 2005
  • Purpose:In an attempt to predict the interpersonal differences of therapeutic response to antipsychotic drugs on pharmaco-genetic bases, this study was designed to investigate the relationship between the therapeutic response to antipsychotic drugs and Taq I A dopamine $D_2$ receptor polymorphism in schizophrenic patients. Methods:The subjects were 158 patients diagnosed with schizophrenia(DSM-IV). The therapeutic response to antipsychotic drugs was evaluated using the Treatment Response Scale(TRS) retrospectively. Patients were divided into two groups, dopamine receptor antagonist responders, and serotonin-dopamine antagonist responders. The patients' Taq I A dopamine $D_2$ receptor polymorphism was determined by polymerase chain reaction(PCR) and restriction fragment length polymorphism(RFLP). Results:The dopamine receptor antagonist responders had the A1 allele in significantly higher incidences (${\chi}^2$(1)=4.875, p=0.027, two-tailed). No significant difference was found among the serotonin-dopamine antagonist responders between those with or without the A1 allele. Conclusions:The patients with the A1 allele responded better to dopamine receptor antagonists than those with no A1 allele. Based on these results, it is suggested that the pharmacological effect of dopamine receptor antagonists can be predicted depending on the presence of the A1 allele in schizophrenic patients.

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Novel SNP in the coding region of the FTO gene is associated with marbling score in Hanwoo (Korean cattle)

  • Chung, Eui-Ryong
    • Journal of Animal Science and Technology
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    • v.56 no.8
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    • pp.27.1-27.6
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    • 2014
  • The fat mass and obesity associated (FTO) gene plays an important role in the regulation of energy homeostasis, fat deposition and obesity. For this reason, the FTO gene is a physiological and functional candidate gene for carcass and meat quality traits in beef cattle. The objectives of this study were to identify SNPs in the exonic regions of FTO gene and to evaluate the association of these SNPs with carcass traits in Hanwoo (Korean cattle). In this study, we newly identified two exonic SNPs in Hanwoo population. The g.125550A > T SNP was located in exon 3 and the g.175675C > T SNP was located in exon 6. Genotyping of the two SNP markers was carried out using PCR-RFLP analysis in Hanwoo steers to evaluate their association with carcass traits. As a result, g.125550A > T SNP genotype was significantly associated with effects on marbling score. Animals with the AA and TT homozygous genotypes had a significantly higher marbling score (p < 0.001) than those with AT heterozygous genotype, and this was significant after Bonferroni correction of the significance threshold (p = 0.003). Dominance effect was also observed for the marbling score (P < 0.05) with higher marbling score of homozygous animals. However, no significant associations with meat quality traits were observed for the g.175675C > T SNP. Our results suggest that the exonic SNP g.125550A > T in the FTO gene may be used as a DNA marker for the selection of Hanwoo with higher marbling.

Antipsychotics Induced Etrapyramidal Symptoms in Schizophrenics in Relation to Cytochrome P450 2D6 Genotype (정신분열병 환자에서 Cytochrome P450 2D6 유형에 따른 부작용에 대한 연구)

  • Lee, Min Soo;Nam, Jong Won
    • Korean Journal of Biological Psychiatry
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    • v.6 no.2
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    • pp.189-192
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    • 1999
  • The genetically determined CYP2D6 activity may be considered to be associated with antipsychotic induced extrapyramidal side effects with interindividual variation. Genetic polymorphism of CYP2D6 was determined by polymerase chain reaction(PCR) and MspI restriction fragment length polymorphisms(RFLP) for 194 schizophrenics. Subjects with a 334bp band were classified a1a1, those with 229bp and 105bp bands a2a2, and those with all three bands a1-a2. We did not identify schizophrenic subject with poor metabolizer. 194 schizophrenic patients previously treated neuroleptic medication, were assessed by Extrapyramidal Symptom Rating Scale(ESRS).The cases were composed of 33 akathisia, 47 parkinsonism, 21 tardive dyskinesia. These results are similar to the previous understanding that the poor metabolizer is very rare in Orientals compared to Caucasians, therefore, it considered that CYP2D6 genotypes have maybe no association with schizophrenia and extrapyramidal side effects in Koreans.

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The CDH2 Gene Polymorphism in Schizophrenia (정신분열병의 CDH2 유전자 다형성)

  • Lee, Won Seok;Kim, Mi Kyung;Jung, Han Yong;Woo, Sung Il;Kwon, Young Joon;Kim, Jong Woo;Lee, Hee Je
    • Korean Journal of Biological Psychiatry
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    • v.12 no.1
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    • pp.62-67
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    • 2005
  • Objective:There has been increasing evidence that neurodevelopmental dysfunction is involved in the pathophysiology of schizophrenia. Cadherin is known to be one of the important molecules in neurodevelopment. This study was performed to examine the relationship between T816C polymorphism of CDH2 gene and schizophrenia. Methods:Genoytypes of T816C polymorphism of CDH2 gene were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 156 Korea patents with schizophrenia and 170 controls. Results:No difference was found between the patients with schizophrenia and the controls in genotype and allele frequencies of T816C polymorphism of CDH2 gene. Conclusion:The results of this study do not support an association between T816C polymorphism of CDH2 gene and schizophrenia. However, it is necessary to investigate other polymorphic regions of CDH2 in schizophrenia.

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Differentiation of Phytoplasmas Infecting Zizyphus jujuba and Paulownia coreana Using PCR-RELP

  • Han, Mu-Seok;Noh, Eun-Woon;Yun, Jeong-Koo
    • The Plant Pathology Journal
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    • v.17 no.4
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    • pp.189-193
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    • 2001
  • The relationships between the phytoplasmas infecting Zizyphus jujuba and Paulownia coreana were investigated by PCR-RELP. The 16S rRNA genes of the phytoplasmas were analyzed and compared with each other after PCR amplification. The amplified bands 1.4 kb in size were analyzed by both restriction digestion and sequencing after cloning into a plasmid vector. In some cases, two different kinds of inserts were observed in the isolates that originated from a single plant. However, many of them appeared to be the amplification products of chloroplastic 16S rRNA gene of host plants. The phytoplasma gene could be differentiated from the chloroplastic gene by restriction digestion of the plasmids carrying the amplification products. Only the recombinant plasmids carrying phytoplasma 16S rRNA gene produced a 1.4 kb band when digested with the enzyme BanII. Of the 52 recombinant plasmids analyzed, 42 appeared to contain inserts that originated from the chloroplastic 16S rRNA gene of the host plants. No variation was detected among 16S rRNA gene of nine phytoplasma isolates infecting Z. jujuba. However, the phytoplasmas infecting Z. jujuba were different from that infecting P. coreana.

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PCR-Based Detection of Mycoplasma Species

  • Sung Hyeran;Kang Seung Hye;Bae Yoon Jin;Hong Jin Tae;Chung Youn Bok;Lee Chong-Kil;Song Sukgil
    • Journal of Microbiology
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    • v.44 no.1
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    • pp.42-49
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    • 2006
  • In this study, we describe our newly-developed sensitive two-stage PCR procedure for the detection of 13 common mycoplasmal contaminants (M. arthritidis, M. bovis, M. fermentans, M. genitalium, M. hominis, M. hyorhinis, M. neurolyticum, M. orale, M. pirum, M. pneumoniae, M. pulmonis, M. salivarium, U. urealyticum). For primary amplification, the DNA regions encompassing the 16S and 23S rRNA genes of 13 species were targeted using general mycoplasma primers. The primary PCR products were then subjected to secondary nested PCR, using two different primer pair sets, designed via the multiple alignment of nucleotide sequences obtained from the 13 mycoplasmal species. The nested PCR, which generated DNA fragments of 165-353 bp, was found to be able to detect 1-2 copies of the target DNA, and evidenced no cross-reactivity with the generated DNA of related microorganisms or of human cell lines, thereby confirming the sensitivity and specificity of the primers used. The identification of contaminated species was' achieved via the performance of restriction fragment length polymorphism (RFLP) coupled with Sau3AI digestion. The results obtained in this study furnish evidence suggesting that the employed assay system constitutes an effective tool for the disagnosis of mycoplasmal contamination in cell culture systems.

Investigation of SNPs in FABP3 and FABP4 Genes and Their Possible Relationships with Fatty Acid Composition in Broiler

  • Maharani, Dyah;Park, Hee-Bok;Jung, Yeon-Kook;Jung, Samooel;Jo, Cheo-Run;Lee, Jun-Heon
    • Korean Journal of Poultry Science
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    • v.38 no.3
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    • pp.231-237
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    • 2011
  • There is limited information of the genetic effect for fatty acid composition in chicken meat. This study assessed the association of FABP3 and FABP4 genes affecting fatty acid composition in broilers. Two single nucleotide polymorphisms (SNPs) were detected in FABP3 gene and five SNPs were identified in FABP4 gene. The SNPs located in intron 1 and exon 1 of FABP3 and FABP4, respectively, were used for genotyping using PCR-RFLP method. The SNPg.285C >T in FABP4 showed suggestive association with high arachidonic acid (C20 : 4) in CT genotypes (P = 0.068). However, the SNP g.508C > T in FABP3 showed no significant associations with fatty acid composition. These results are the first report to investigate the SNPs in FABP3 and FABP4 genes and their associations with fatty acid composition, although we only found the possible association of FABP4 SNP with fatty acid composition. These results should provide valuable information for further investigation of the genes affecting fatty acid composition in chicken.

Genetic polymorphism of Estrogen metabolising enzymes and individual genetic susceptibility to breast cancer in Korean (Estrogen대사 효소의 유전자 다형성과 한국인 유방암 환자의 유전적 감수성에 대한 연구)

  • 김현준;이수진;공구
    • Environmental Mutagens and Carcinogens
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    • v.23 no.1
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    • pp.23-29
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    • 2003
  • To determine the frequencies of the genotypes of estrogen metabolising enzyme (CYP17, CYP1A1, CYP1B1, and COMT) and to identify the high-risk genotypes of these metabolic enzymes to breast cancer in Korean, the author has analysed 115 breast cancer patients and corresponding age and sex matched heathy controls using polymerase chain reaction-restiction fragment length polymorphism (PCR-RFLP). A2/A2 genotype in CYP17 polymorphism, m2/m2 genotype in CYP1A1 polymorphism, and Val/Val genotype in CYP1B1 had 0.95, 1.40 and 0.76 relive risks to breast cancer comparing with reference genotypes of each polymorphism, respectively. Among the genotypes of COMT enzyme polymorphism, L/H and L/L genotypes had 0.97 and 1.54 relative risks to breast cancer, respectively. According to the number of high risk genotype, the patients with one or two putative high risk genotypes had 0.95 and 1.94 relative risks to breast cancer, respectively. This study have demonstrated the unique frequency of genotypes of estrogen metabolizing enzyme in Korean healthy women, which will provide the basic data and insights to study the estrogen related conditions in Korean women including breast and endometrial cancers. And it also indicates that the well-known high risk genotypes of estrogen metabolizing enzymes are not significantly associated with the development of breast cancer in Korean women.

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Phylogenetic Analysis of Phenanthrene-Degrading Sphingomonas

  • Han, Kyu-Dong;Jung, Yong-Tae;Son, Seung-Yeol
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.942-948
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    • 2003
  • Soil samples were obtained from 5 sites contaminated with polycyclic aromatic hydrocarbons (PAHs). These soil samples were cultured in using phenanthrene as a sole carbon and energy source, and 36 strains of phenanthrene-degrading bacteria were isolated from 3 sites. Most of them degraded 500 ppm of phenanthrene within 8 to 10 days, and these isolates could degrade a few other PAHs other than phenanthrene. Their genotypes were determined by restriction digests of the l6S rRNA genes [amplified ribosomal DNA restriction analysis (ARDRA)]. It was found that all the phenanthrene degrading isolates were included in 4 ARDRA types, and they showed a strict site endemism. l6S rDNAs of 12 strains selected from different sites were sequenced, and they were all confirmed as Sphingomonas strains. Their l6S rDNA sequences were compared for phylogenetic analysis; their sequence showed a similar result to ARDRA typing, thus indicating that these heterotrophic soil bacteria are not regionally mixed. In addition, it was found that the microbial diversity among sampling sites could be monitored by l6S rDNA PCR-RFLP pattern alone, which is simpler and easier to perform, without l6S rDNA sequence analysis.