• Parasites, Hosts and Diseases
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• v.53 no.3
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• pp.271-277
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• 2015

The genetic diversity of Schistosoma haematobium remains largely unstudied in comparison to that of Schistosoma mansoni. To characterize the extent of genetic diversity in S. haematobium among its definitive host (humans), we collected S. haematobium eggs from the urine of 73 infected schoolchildren at 5 primary schools in White Nile State, Sudan, and then performed a randomly amplified polymorphic DNA marker ITS2 by PCR-RFLP analysis. Among 73 S. haematobium egg-positive cases, 13 were selected based on the presence of the S. haematobium satellite markers A4 and B2 in their genomic DNA, and used for RFLP analysis. The 13 samples were subjected to an RFLP analysis of the S. haematobium ITS2 region; however, there was no variation in size among the fragments. Compared to the ITS2 sequences obtained for S. haematobium from Kenya, the nucleotide sequences of the ITS2 regions of S. haematobium from 4 areas in Sudan were consistent with those from Kenya (> 99%). In this study, we demonstrate for the first time that most of the S. haematobium population in Sudan consists of a pan-African S. haematobium genotype; however, we also report the discovery of Kenyan strain inflow into White Nile, Sudan.

• Interdisciplinary Bio Central
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• v.4 no.3
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• pp.8.1-8.7
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• 2012

The motor neuron degeneration 2 (mnd2) mice carry a point mutation of A to T nucleotide transversion at the serine 276 residue of high temperature requirement A2 (HtrA2), resulting in losses of an AluI restriction enzyme site (5'AGCT3') and the HtrA2 serine protease activity. Moreover, dysfunctions of HtrA2 are known to be intimately associated with the pathogenesis of neurodegenerative diseases, including Parkinson's disease. Thus, this mnd2 mouse is an invaluable model for understanding the physiological role of HtrA2 and its pathological role in neurodegenerative diseases. Nevertheless, many molecular and cellular biologists in this field have limited experience in working with mutant mouse models due to the necessity of acquired years of the special techniques and knowledges. Herein, using the mnd2 mouse model as an example, we describe easy-to-use standard protocols for web-based analyses of target genes, such as HtrA2, and a novel approach for simple and accurate PCR-AluI-RFLP-based genotype analysis of mnd2 mice. In addition, band resolution of AluI-RFLP fragments was improved in 12% polyacrylamide gel running in 1X Tris-Glycine SDS buffer. Our study indicates that this PCR-AluI-RFLP genotype analysis method can be easily applied by the molecular and cellular biologist to conduct biomedical science studies using the other mutant mouse models.

• Korean Journal of Soil Science and Fertilizer
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• v.41 no.6
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• pp.415-418
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• 2008

The application of sludge wastes into agricultural fields has been increasing annually in Korea. In particular, sewage sludge application has been widely accepted in decades. Sewage sludge application aid in the recycling of essential nutrients and act as a source of organic matter improving the structure and water-holding properties of the soil. The efficient use of sludge wastes, however, requires an individual assessment of waste products. This study assessed the biological characteristics of organic waste-treated lysimeter soils and develop its indicator to assess the soil health of organic waste-treated lysimeter soils. Several analytical techniques more recently developed such as restriction fragment length polymorphism (RFLP), phospholipid fatty acid (PLFA), and community level substrate utilization (CLSU) fingerprints allow for detailed analyses of soil microbial communities. PLFA and RFLP was, therefore, used in the study to characterize the microbial communities in soil without the need to isolate individual fungi and bacteria. PLFA, RFLP and CLSU have been utilized to assess microbial characteristics of the lysimeter soils with four different sludge wastes for eight consecutive years. Each of these methods was analyzed for a different aspect of soil microbial characteristics. The study would disclose those methods yielded highly reproductive results for each soil and allow distinguishing the soils based on the structures of specific geneand PLFA-pools more than CLSU fingerprints. PLFA methods, especially, revealed the same relative similarities of the treated soils based on cluster analysis of the biological characteristics. Pig manure compost-treated soil, however, was only the same relative resemblance among the three methods. These results indicated that PLFA easily assessed the biological soil characterization.

• The Korean Journal of Mycology
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• v.28 no.2
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• pp.112-117
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• 2000

A similarity coefficient were analyzed by RFLP of fourteen species of entomopathogenic fungi, isolated from inhabiting pupa and adult insect at forest. Each rDNA ITS I and ITS II with primers of ITS 1 and ITS 4 was amplified by PCR. The amplified products were conserved to 500 bp were not demarcated between genus and species. Four Paeciliomyces tenuipes, two Beauveria bassiana and six Cordyceps militaris were treated by seven restriction enzymes and confirmed in species except JB3 by electrophoresis band. However, the band of C. scarabaeicola showed the identity with B. bassiana. The result of this experiment indicated that the teleomorph of C. scarabaeicola was the same as that of B. bassiana. CfoI and HpaII of seven restricted enzymes were easily discriminating in the genus between Paecilomyees and Cordyceps. Especially, CfoI was more effective to classify the genera of Paecilomyees, Cordyceps and Beauveria than other restriction enzymes. The band patterns of RFLP of P. tenuipes, C. militaris, C. scarabaeicola and B. bassiana were also analyzed by UPGMA program of NTSYS-pc and showed 100% significance. Thus, the similarity coefficient tended to be lower between genera by RFLP analysis, but was higher between species.

• Journal of Microbiology
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• v.34 no.4
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• pp.295-299
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• 1996

To infer phylogenetic relationships between species of Trichaptum (Polyporaceae), RFLP analyses of PCR-amplified DNAs were accomplished. Regions coding for ITSs of nuclear SSU rRNA genes and for mitochondrial SSU rRNA genes from thirteen strains of four Trichaptum species (T. abietinum, T. biforme, T. fusco-violaceum, and T. laricinum) were amplified and digested with eight restriction enzymes. All the fragmentation patterns were characterized and coded as 0/1 for the absence/presence of fragments. A phylogenetic tree based on the combined data sets was constructed using the Dollo parsimony method. While every two strains of T. abietinum, T. biforme, T. fusco-violaceum, and T. laricinum formed an independent group, the other strains of T. abietimum and T. fusco-violaceum made mixed groupings among compared strains. It is inferred that T. abietinum and T. fusco-violaceum have more variations, possibly geographic or physiological ones, than other species in the genus.

• Journal of Microbiology and Biotechnology
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• v.22 no.4
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• pp.563-566
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• 2012

To investigate the possibility of horizontal gene transfer between agricultural microorganisms and soil microorganisms in the environment, Bacillus subtilis KB producing iturin and the PGPR recombinant strain Pseudomonas fluorescens MX1 were used as model microorganisms. The soil samples of cucumber or tomato plants cultivated in pots and the greenhouse for a six month period were investigated by PCR, real-time PCR, Southern hybridization, and terminal restriction fragment length polymorphism (T-RFLP) fingerprinting. Our data from Southern blotting and T-RFLP patterns suggest that the model bacteria do not give significant impacts on the other bacteria in the pots and greenhouse during cultivation.

• Korean Journal of Pharmacognosy
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• v.28 no.1
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• pp.1-8
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• 1997

Bangpoong (防風) is a popular crude drug used to expel wind from the body surface (祛風解表) to remove dampness (勝濕). And to relieve pain (正痛) and spasm (正痙). In China and Japan, roots of Saposhnikovia divaricata (Turcz.) Schischk. Is used as Bangpoong. However, the roots of Peucedanum japonicum Thunb. Or Glehnia littoralis (A. Gray) Fr. Schmidt ex Miquel, being called Sikbangpoong (植防風) and Wonbangpoong (元防風) respectively are used instead of Bangpoong in Korea. The ITS regions of nuclear ribosomal DNA were analyzed to determine original plants and to design a molecular identification method for the crude drugs used as Bangpoong in Korea and China. It is demonstrated that RFLP analysis via PCR has the great potential as a novel tool to test crude drugs for the quality control and standardization.

• Mycobiology
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• v.30 no.1
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• pp.1-4
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• 2002

Genetic relatedness of medically important Exophiala species such as E. dermatitidis, E. mansonii, and three E. jeanselmei varieties: jeanselmei, lecanii-corni, and heteromorpha was examined using PCR-RFLP(restriction fragment length polymorphism) of ribosomal DNA, M-13, $(GTG)_5$ and nucleotide sequences of ribosomal ITS(internal transcribed space) II regions. Three E. jeanselmei varieties showing distinct band patterns for each DNA markers as well as different nucleotide sequences of ribosomal ITS II regions could be considered as a separate species. E. dermatitidis and E. mansonii demonstrated the identical band patterns of RFLP of ribosomal DNA, M-13, and $(GTG)_5$ markers. However, nucleotides sequences of ribosomal ITS II region were different between these two species.

• Proceedings of the Plant Resources Society of Korea Conference
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• v.18 no.1
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• pp.17-25
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• 2005

• 한국약용작물학회:학술대회논문집
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• 2009.05a
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• pp.415-416
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• 2009