This study was carried out to evaluate the suitability of microsatellite markers for varietal identification and genetic relationship of 170 commercial pepper cultivars. The relationship between marker genotypes and 11 pepper cultivars with different morphological traits was also analyzed. Of the 302 pairs of microsatellite primers screened against 11 pepper cultivars, 24 pairs were highly polymorphic in terms of number of alleles. These markers were applied for the construction of DNA profile data base for 170 commercial pepper cultivars. A total of 164 polymorphic amplified fragments were obtained from 24 microsatellite primers. The average polymorphism information content was 0.673 ranging from 0.324 to 0.824. One hundred and sixty four microsatellite alleles were used to calculate Jaccard's distance coefficients using unweighted pair group method. A clustering group of varieties, based on the results of microsatellite analysis, were categorized into 3 major groups corresponding to morphological traits. The phenogram discriminated all varieties by markers genotypes. These microsatellite markers will be useful as a tool for protection of plant breeders' intellectual property rights through variety identification in distinctness, uniformity and stability test.
Kim, Ho-Joong;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Kim, Keun-Youl;Han, Yong-Chol
Tuberculosis and Respiratory Diseases
/
v.40
no.5
/
pp.509-518
/
1993
Background: By amplifying small amount of DNA, polymerase chain reaction (PCR) can be used for the detection of very small amount of microbial agent, and may be especially useful in certain cases which are difficult to be diagnosed microbiologically or serologically. Tuberculous pleurisy is a disease that can be diagnosed in only 70% of cases by conventional diagnostic tools, and PCR would be a very rapid, easy, and sensitive diagnostic method. Method: The specificity and sensitivity of PCR to detect Mycobacterium tuberculosis DNA were evaluated using various strains of Mycobacteria. To evaluate the diagnostic usefulness of PCR in tuberculous pleurisy, we used PCR to detect Mycobacterium tuberculosis DNA in pleural fluid. The amplification target was 123 base pair DNA, a part of IS6110 fragment, 10~16 copies of which are known to exist per genome. The diagnostic yield of PCR was compared with conventional methods, including pleural fluid adenosine deaminase (ADA) activity. Also, the significance of PCR in undiagnosed pleural effusion was evaluated prospectively with antituberculosis treatment. Results: 1) Using cultured Mycobacterium tuberculosis and other strains, PCR could detect upto 1 fg DNA and specific for only Mycobacterium tuberculosis and Mycobacterium bovis. 2) Using pleural effusions of proven tuberculosis cases, the sensitivity of PCR was 80.0% (16/20), and the specificity 95.0% (19/20). 3) Among 13 undiagnosed, but suspected tuberculous effusion, the positive rate was 60% in 10 improved cases after antituberculosis medications, and 0% in 3 cases of proven malignancy later. 4) Adenosine deaminase level of proven and clinically diagnosed tuberculous pleurisy patients was significantly higher than that of excluded patients, and correlated well with PCR results. Conclusion: We can conclude that PCR detection of Mycobacterium tuberculosis in pleural effusion has acceptable sensitivity and specificity, and could be an additional diagnostic tool for the diagnosis of tuberculous pleurisy.
Objective: Mongolia is one of a few countries that supports over 25 million goats, but genetic diversity, demographic history, and the origin of goat populations in Mongolia have not been well studied. This study was conducted to assess the genetic diversity, phylogenetic status and population structure of Mongolian native goats, as well as to discuss their origin together with other foreign breeds from different countries using hypervariable region 1 (HV1) in mtDNA. Methods: In this study, we examined the genetic diversity and phylogenetic status of Mongolian native goat populations using a 452 base-pair long fragment of HVI of mitochondrial DNA from 174 individuals representing 12 populations. In addition, 329 previously published reference sequences from different regions were included in our phylogenetic analyses. Results: Investigated native Mongolian goats displayed relatively high genetic diversities. After sequencing, we found a total of 109 polymorphic sites that defined 137 haplotypes among investigated populations. Of these, haplotype and nucleotide diversities of Mongolian goats were calculated as 0.997±0.001 and 0.0283±0.002, respectively. These haplotypes clearly clustered into four haplogroups (A, B, C, and D), with the predominance of haplogroup A (90.8%). Estimates of pairwise differences (Fst) and the analysis of molecular variance values among goat populations in Mongolia showed low genetic differentiation and weak geographical structure. In addition, Kazakh, Chinese (from Huanghuai and Leizhou), and Arabian (Turkish and Baladi breeds) goats had smaller genetic differentiation compared to Mongolian goats. Conclusion: In summary, we report novel information regarding genetic diversity, population structure, and origin of Mongolian goats. The findings obtained from this study reveal that abundant haplogroups (A to D) occur in goat populations in Mongolia, with high levels of haplotype and nucleotide diversity.
The study, using the input-output analysis of 402 industrial sectors by Bank of Korea(BOK) and the resulting outcomes of price model, aims to evaluate the spillover effects the international fluctuations in crude oil prices have on the commodities prices and consequently, analyse the management and profitability of the offshore fisheries in Korea. At present, the fisher men are provided with tax-free oils for their fishing operations as specified under the Special Tax Treatment Control Law. However, the exhaustion of marine resources and new international fisheries agreements, which resulted in the loss of fishing grounds, made the stable catch even more unpredictable and the hike in the price of the international crude oil would have adverse effects on the fishing industry. The study revealed that the increasing rise in the price of crude oil would exert sweeping spillover effects on other industry sectors in general and accordingly, lead to a poorer performance by fisheries. The price spillover coefficients for the diesel oil was 0.6026, which would translate into the 42.6% increase in the prices of oil when the increase ratio of 73.3% for the base crude oil was applied based on the calculation methods employed in the study. This in turn increased the ratio of diesel oil required in the offshore fisheries from 23.3% to 16.6%, diminishing the ratio of current net profits to minus 2.0% from 4.2% otherwise. By fishing type, the Pair Trawl suffered current net profits loss most by ratio of minus 9.4% and other fisheries such as Coastal Stow Nets, Coastal Angling, Danish Sein also suffered ratio of 7% and more in the loss of current net profits. With the deteriorating fishing performance, coupled with the increasing international crude oil prices, it is urgently required that the authorities concerned deliberate in depth on such schemes as follows in efforts to secure stable fishing production. First, provision of large-scale storage facilities for oil is needed to timely adapt to the fluctuations in international crude oil prices. Secondly, in line with the stabilization of tax-free oil prices, duty levied on oils for fishing and tax collected from the refineries need to be tax-exempt. Thirdly, the beneficiaries from the provision of tax-free oil should be broadened, not limited to special fishing operation only. Fourth, investment in stabilization of the oil prices should be encouraged, possibly through funding from the formation of fisheries development funds underway.
The genetic structures at one leucine aminopeptidase locus (LAP-A) of acorns, seedling raised in greenhouse and forest form the two beech provenances. West Germany and Rumania, were investigated and compared with each other. In many pair wise comparisons significant differences in genotypic structure as well as genic structure were ascertained between different developmental stages. In both the provenances, the allele $A_2$ seems to have advantage at both seedling stages raised under two different conditions. Homozygous carriers of $A_2$ allele survived best in greenhouse, while heterozygous carriers especially with $A_2$ allele possessed great viability under more variable environmental conditions. Since a distinct different genetic background was present in two base populations, the identical effect of the allele $A_2$ confirms the adaptiveness of this locus. With aid of some measures such as viability parameter and genetic distance, the character of occurred viability selection of further explained. The possible significance of this locus at this early stage is discussed in relation to adaptation of this long lived tree species to heterogeneous environment.
Viroids are about 250-400 base pair of short single strand RNA fragments have been associated with economically important plant diseases. Due to the lack of protein expression capacity associated with replication, it is very difficult to diagnosis viroid diseases in serological methods. For detecting viroid at plants, molecular-based techniques such as agarose gel electrophoresis, polyacrylamide gel electrophoresis (PAGE), DNA-hybridization, blotting analysis and conventional RT-PCR are reliable. Real-time RT-PCR methods that grafted on RT-PCR methods with improved confirmation methods have been also utilized. However, they are still labor-intensive, time-consuming, and require personnel with expertise. Loop-mediated Isothermal Amplification (LAMP) method is a nucleic acid amplification method under the isothermal condition. The LAMP methodology has been reported to be simple, rapid, sensitive and field applicable in detecting a variety of pathogens. The results of LAMP method can be colorized by adding a visible material such as SYBR green I, Evagreen, Calcein, Berberine and Hydroxy naphthol blue (HNB) with simple equipment or naked eyes. The combination of LAMP method and nucleic pathogens, viroids, can be used to realize simple diagnosis platform for the genetic point-of care testing system. The aim at this review is to summary viroid-caused diseases and the simple visible approach for diagnosing viroids using Loop-mediated Isothermal Amplification (LAMP) method.
Objects : Clozapine, prototype of the atypical neuroleptics, was known to have unique antipsychotic effect with a few extrapyramidal effects. While most typical antipsychotic agents mainly block $D_2$ receptors, clozapine has higher affinity for dopamine $D_4$ receptor than for $D_2$ receptor. Many researchers have tried to find out the relationship between schizophrenia and the abnormality of the genes coding dopamine receptors. But no consistent findings were reported. Recently, dopamine $D_4$ receptor was fully sequenced, and the alleles of dopamine $D_4$ receptor gene was found in unusual form on the 48th base pair. Our study was performed to identify the distribution of the dopamine $D_4$ receptor alleles of schizophrenics and normal controls, and whether any difference between the dopamine $D_4$ receptor alleles of schizophrenics and that of normal controls exists. Methods : DNA was extracted from the blood of schizophrenic patients(N=60) and normal controls(N=60). Part of the dopamine $D_4$ receptor gene was amplified by PCR, and amplified DNA was electrophoresed. Authors compared the distribution of the alleles of dopamine $D_4$ receptor gene of normal controls and that of schizophrenic patients. Results : Six kinds of alleles of $D_4$ receptor were observed both groups. The fourth repeat form of alleles was the most common in both schizophrenic patients(75.8%) and normal controls(70.3%), so there was not significant difference between two groups. Conclusion : The Difference in the distribution of the dopamine $D_4$ receptor gene alleles is not thought to be responsible for the pathophysiology of the schizophrenia. However, the difference in the expression of the dopamine $D_4$ receptor gene between normal and schizophrenia is left to be scrutinized.
Trichophyton erinacei is a dermatophyte pathogen that infects both humans and hedgehogs. A two-month old female four-toed hedgehog presented to the Chonbuk Animal Medical Center with pruritus, excoriation and crust on her face for ten days. The owner of the hedgehog also exhibited the clinical signs of scaly erythema with fine vesicles on her neck. A presumptive diagnosis of dermatophytosis was made based on the results of an acetate tape preparation in which hyphae and chains of arthroconidia were observed. The crusts from the lesions were then cultured on Sabouraud Dextrose Agar for identification. After 10 days of incubation, downy colored colonies that had a central umbo with a white granular surface and a yellow pigment ring in the reverse were observed. Microscopic analysis revealed the presence of numerous teardrop shaped microconidia singly attached to the sides of the hyphae. In addition, 2-6 roomed macroconidia that were somewhat irregular in shape and size were present, and abundant intermediate sized spores were observed between the micro and macro conidia. To confirm that the culture was T. erinacei, the internal transcribed spacer region of the 5.8S phase of the ribosomal RNA gene (ITS1-5.8S-ITS2 rDNA) was amplified by PCR and then sequenced. A 679-base pair fragment of DNA was then compared with sequences in GenBank and found to be 99% homologous with sequences of T. erinacei (Z97997 and Z97996. The clinical signs were resolved after four weeks of treatment with oral and topical ketoconazole and chlorhexidine. To the best of our knowledge, this represents the first case of T. erinacei isolated from a four-toed hedgehog in Korea.
Leucine-rich repeat containing protein 10 (LRRC10) is characterized as a cardiac-specific gene, suggesting a role in heart development and disease. A severe cardiac morphogenic defect in zebrafish morphants was recently reported but a contradictory result was found in mice, suggesting a more complicated molecular mechanism exists during mouse embryonic development. To elucidate how LRRC10 is regulated, we analyzed the 5'enhancer region approximately 3 kilo bases (kb) upstream of the Lrrc10 start site using luciferase reporter gene assays. Our characterization of the Lrrc10 promoter indicates it possesses complicated cis-and trans-acting elements. We show that GATA4 and MEF2C could both increase transcriptional activity of Lrrc10 promoter individually but that they do not act synergistically, suggesting that there exists a more complex regulation pattern. Surprisingly, knockout of Gata4 and Mef2c binding sites in the 5’enhancer region (-2,894/-2,889) didn't change the transcriptional activity of the Lrrc10 promoter and the likely GATA4 binding site identified was located in a region only 100 base pair (bp) upstream of the promoter. Our data provides insight into the molecular regulation of Lrrc10 expression, which probably also contributes to its tissue-specific expression.
The rapid and reliable 16S rDNA multiplex polymerase chain reaction (PCR) assay was established to characterize bacterial etiologies of middle ear effusion. These etiologies included Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pneumonia, which were detected in middle-ear effusion (MEE) samples taken from patient with otitis media. A total of 39 MEE samples were aspirated from 26 patients. DNA was extracted from MEE samples, and PCR was done with DNA extracts by using the common primers, which is localized at C4 region in the 16S rDNA gene of all bacterial species, and species-specific primers: (i) Haemophilus-specific primer, (ii) Moraxella- specific primer, and (iii) Streptococcus-specific primer. Among 39 samples tested, 24 (61.5%) were positive for H. influenzae, 10 (25.6%) were positive for M. catarrhalis, 3(7.7%) were positive for S. pneumonia, and 11 (28%) were negative for 165 rDNA multiplex PCR reaction. Nine samples (28.6%) exhibited a mixed infection and were positive for both H. infuenzae and M. catarrhalis. We suggested that 16S rDNA multiplex PCR is a useful method to identify rapidly for rapid identification of the pathogenic bacteria and characterization of bacterial etiologies of middle ear effusion.
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