• Horticultural Science & Technology
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• v.35 no.1
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• pp.98-107
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• 2017

A DNA profile database was constructed to investigate the genetic relatedness of 72 germplasm samples of Pyrus and related cultivars using microsatellite markers. Three P. pyrifolia, four P. commus, and one P. betulifolia cultivars with different morphological traits were screened using 387 pairs of microsatellite primers. A core set of 11 primer pairs was selected to obtain 133 polymorphic amplified fragments meeting three criteria: high polymorphism information contents (PIC), high repeatability, and distinct allele patterns. The number of alleles per locus ranged between 4 and 22. Average PIC was 0.743 (range: 0.557 - 0.879). Cluster analysis using the unweighted pair - group method with arithmetical average (UPGMA) separated the 72 pear cultivars and germplasm samples into four major groups: Chinese, European pears, and a cluster of 55 Asian pears that could be reclassify into two subcluster, I - $1^{st}$ and II - $2^{nd}$, according to pedigree information. Almost all of the cultivars were discriminated by 11 microsatellite marker genotypes. The microsatellite DNA profile database may be utilized as tool to verify distinctness, uniformity, and stability between candidate cultivar, and to verify in the distinctness of existing cultivars.

• Journal of Pharmacopuncture
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• v.3 no.1
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• pp.35-51
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• 2000

The purpose of this study is to prove the analgesic effects of apitoxin and its mechanism via jaw-opening reflex(JOR) and measuring expression of mRNA in Phospholipase and Tryptophan hydroxylase(TPH) using RT-PCR. The experiments were carried out on Sprague-Dawley rats(300-400g) and mastocytoma(P-185 HTR) for JOR and RT-PCR, respectively. Rats anesthetized with thiopental sodium (80mg/kg) were used in the Tooth Pulp stimulation induced JOR. The amplitude of a digastric electromyogram (dEMG) was recorded during the stimulation at an intensity of 1.5 times the threshold for JOR. Apitoxin used in this experiment was diluted with normal saline by 1:1000. Apitoxin was injected intravenously into the test group while normal saline to the control group. However, it was injected directly into the cell of mastocytoma. We referred to base sequence registered in Genbank in designing primers for RT-PCR. The results were as follows; (1)Compared with control group, analgesic effect started to show right after Sprague-Dawely rats were treated with apitoxin($71.50{\pm}8.08$) and lasted for 50 minutes. (2)As a result of the experiment of RT-PCR, we witnessed significant changes in the degree of expression of phospholipase or rate-limiting enzyme of biosynthesis of prostaglandins with $10{\mu}g/ml$ apitoxin.($31.74{\pm}18.98%$, P<0.05) (3)As a result of the experiment of RT-PCR, we witnessed significant changes in the degree of expression of TPH or rate-limiting enzyme in biosynthesis of serotonin with $10{\mu}g/ml$ apitoxin.($131.37{\pm}16.87%$, P<0.05). These results suggest that $10{\mu}g/ml$ apitoxin have the most analgesic effects. This study showed that apitoxin has analgesic effects and held good for 50 minutes. The injection of apitoxin has brought out changes in the degree of expression of phospholipase and TPH. These results strongly suggest that analgesic mechanism by apitoxin is closely related to prostaglandins and serotonin.

• Journal of Mushroom
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• v.9 no.2
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• pp.59-62
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• 2011

To develop a new cultivar of King oyster mushroom(Pleurotus eryngii), G09-21 as parental strain was selected by the method of Di-mon crossing between monokaryotic strains derived from ASI 2824(Keunneutari No.2) and dikaryotic strain ASI 2887(Aeryni 3). The Pe21-51($G09-21-10{\times}2844-11$) was shown the best cultural characteristics, selected to be a new cultivar and named as 'Song-A'. The 'Song-A' was formed incompatibility line distinctly in the confrontation growth of parental strains Keunneutari No.2, Aeryni 3 and ASI 2844. The optimum temperature for mycelial growth, fruiting body development and pH arrange were $25{\sim}30^{\circ}C$, $14{\sim}16^{\circ}C$ and pH5~8, respectively. Fruiting body production per bottle was about $94.7{\pm}29.5$ g which is almost 106% quantity compared to that of other cultuvar Keunneutari No.2. And also the stip is thick and long but the number of available stipe is few. Analysis of the genetic characteristics of the new cultivar 'Song-A' showed a different DNA profile as that of the control strains, Keunneutari No.2, Aeryni 3 and ASI 2844, when RAPD(Random Amplified Polymorphic DNA) primers URP4 and 7 were used. This new cultivar 'Song-A' of Pleurotus eryngii is characterized by a small number of primordia formation and the stip is thick and long. Therefore, we expect that this new strain will save of labor and cost by without culling work.

• Journal of Animal Science and Technology
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• v.58 no.4
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• pp.15.1-15.6
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• 2016

Background: Our previous study had identified the SNP (g.81966377T > C) and indel (g.81966364D > I) located in the promoter of APM1 to have a significant effect on marbling in Hanwoo. APM1 encodes an adipocytokine called adiponectin, which plays a significant role in lipogenesis. The aim of this study was to verify and validate the effect of the SNP and indel on marbling and other carcass traits in a large, representative, countrywide population of Hanwoo cattle. The carcass traits measured were marbling (MAR), backfat thickness (BFT), loin eye area (LEA), and carcass weight (CAW). Results: Primers were designed to amplify 346 bp of the genomic segment that contained the targeted SNP (g.81966377) and the indel (g.81966364). After data curation, the genotypes of 8,378 individuals identified using direct sequencing analysis estimated frequencies for C (0.686) and T (0.314) respectively showing genotype frequencies for CC (0.470), CT (0.430) and TT (0.098). The genotypes were significantly associated with MAR, BFT and LEA. The indel had significant effect on marbling (P < .0001) with strong additive genetic effects. The allele frequencies was estimated at (DEL, 0.864) and insertion (INS, 0.136) presenting genotypes of D/D (75.63 %), D/I (21.44 %), and I/I (2.92 %). Significant departure from Hardy-Weinberg equilibrium was not detected for both the SNP and the indel. Conclusion: The SNP genotypes showed significant association with MAR, BFT and LEA with strong additive genetic effects, while the indel was significantly associated with MAR. The results confirmed that the variants can be used as a genetic marker for improving marbling in Hanwoo.

• Tuberculosis and Respiratory Diseases
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• v.70 no.4
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• pp.315-322
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• 2011

Background: A variety of diagnostic modalities for lung cancer have been developed. To achieve efficient and early detection of lung cancer, we tried to measure the expression rates of the melanoma associated gene (MAGE) and synovial sarcoma on X chromosome (SSX) genes. Methods: We designed primers for the SSX gene. In addition to the pre-developed MAGE A primer, using an SSX gene primer was attempted to increase the detection rate. We obtained cancer tissues and cancer-free lung tissues from resected lung, sputum from lung cancer patients who had not undergone surgery, and sputum from healthy people and patients with benign intrathoracic diseases. Results: The sensitivity of the MAGE or SSX gene RT-PCR to identifying cancer tissue of the 69 lung cancer patients was 95.2% for squamous cell carcinoma (scc), 87.0% for adenocarcinoma, and 100% for small cell carcinoma. The mean sensitivity value was 94.2% (p=0.001). For adenocarcinoma, the additional use of the SSX gene resulted in a higher expression rate than MAGE alone (87% vs. 69.6%). The expression rate for the cancer-free lung tissue was 14.3% in scc, 17.4% in adenocarcinoma, and 25.0% in small cell carcinoma. In the induced sputum of 49 lung cancer patients who had not undergone surgery, the expression rate for one of the two genes was 65.5%. The expression rate for the sputum of healthy people and benign intrathoracic diseases by MAGE or SSX gene reverse transcription polymerase chain reaction (RT-PCR) was 3.8% and 17.7%. Conclusion: Detecting lung cancer using the expression of MAGE and SSX genes in lung cancer tissue has high sensitivity.

• The Korean Journal of Mycology
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• v.36 no.2
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• pp.130-137
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• 2008

To distinct the commercial strains in Pleurotus spp., 81 strains in eight Pleurotus species were used. DNA fingerprinting using URP-PCR was conducted to determine the phylogenetic relationships among Pleurotus strains. DNA profiles of Pleurotus species obtained by twelve URP primers were analyzed for genetic similarity by NTSYS program. We could divide strains into ten clusters, in which three of them belong to P. ostreatus and the others to the different species, respectively. At the 76% similarity level, 70 P. ostreatus strains were distinguished into three clusters. Cluster I contained 35 strains and some of them showed almost 100% similarity, one strain closely related to Weonhyeong and six strains closely related to Wangheukpyeong. In cluster II, twenty-one out of 23 strains showed 100% to Suhan. Cluster III contained twelve strains, including six strains closely related to Chunchu-2. The results suggested that there are many same strains with different names in mushroom spawn market.

• Restorative Dentistry and Endodontics
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• v.20 no.2
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• pp.699-720
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• 1995

This study was conducted to evaluate the effect of benzalkonium chloride solution as a wetting agent instead of water on dentin bonding with NTG-GMA/BPDM system (All-bond 2, Bisco.) and DSDM system (Aelitebond, Bisco.). Benzalkonium chloride solution is a chemical disinfectant widely used in medical and dental clinics for preoperative preparation of skin and mucosa due to its strong effect of cationic surface active detergent. Eighty freshly extracted bovine lower incisor were grinded labially to expose flat dentin surface, and then were acid-etched with 10 % phosphoric acid for 15 second, water-rinsed, and dried for 10 second with air syringe. The specimens were randomly divided into 8 groups of 10 teeth. The specimens of control group were remoistured with water and the specimens of experimental groups were remoistured with 0.1 %, 0.5 %, and 1.0 % benzalkonium chloride solution respectively. And then, the Aelitefil composite resin was bonded to the pretreated surface of the specimens by use of All-bond 2 dentin bonding system or Aelitebond dentin bonding system in equal number of the specimens. The bonded specimens were stored in $37^{\circ}C$ distilled water for 24 hours, then the tensile bond strength was measured, the mode of failure was observed, the fractured dentin surface were examined under scanning electron microscopy, and FT-IR spectroscopy was taken for the purpose of investigating the changes of the dentin surface pretreated with benzal konium chloride solution followed by each primer of the dentin bonding systems. The results were as follows : In the group of bonding with NTG-GMA/BPDM dentin bonding agent(All-bond 2), higher tensile bond strength was only seen in the experimental group remoistured with 0.1 % benzal konium chloride solution than that in water-remoistured control group(p<0.05). In the group of bonding with DSDM dentin bonding agent (Aelitebond), no significant differences were seen between the control and each one of the experimental group(p<0.05). Higher tensile bond strength were seen in NTG-GMAIBPDM dentin bonding agent group than in DSDM dentin bonding agent group regardless of remoistur ization with benzal konium chloride solution. On the examination of failure mode, cohesive and mixed failure were predominantly seen in the group of bonding with NTG-GMAIBPDM dentin bonding agent, while adhesive failure was predominantly seen in the group of bonding with DSDM dentin bonding agent. On SEM examination of fractured surfaces, no differences of findings of primed dentin surface between the groups with and without remoisturization with benzal konium chloride solution. FT-IR spectroscopy taken from the control and the experimental group reve::.led that some higher absorbance derived from the primers binding to dentin surface was seen at the group pretreated with 0.1 % benzal konium chloride solution than at the control group of remoisturizing with water.

• Genes and Genomics
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• v.40 no.12
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• pp.1319-1329
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• 2018

SSRs were successfully isolated from the Perilla crop in our current study, and used to analyze Perilla accessions from East Asia. Analyses of the clear genetic diversity and relationship for Perilla crop still remain insufficient. In this study, 40 new simple sequence repeat (SSR) primer sets were developed from RNA sequences using transcriptome analysis. These new SSR markers were applied to analyze the diversity, relationships, and population structure among 35 accessions of the two cultivated types of Perilla crop and their weedy types. A total of 220 alleles were identified at all loci, with an average of 5.5 alleles per locus and a range between 2 and 10 alleles per locus. The MAF (major allele frequency) per locus varied from 0.229 to 0.943, with an average of 0.466. The average polymorphic information content (PIC) value was 0.603, ranging from 0.102 to 0.837. The genetic diversity (GD) ranged from 0.108 to 0.854, with an average of 0.654. Based on population structure analysis, all accessions were divided into three groups: Group I, Group II and the admixed group. This study demonstrated the utility of new SSR analysis for the study of genetic diversity and population structure among 35 Perilla accessions. The GD of each locus for accessions of cultivated var. frutescens, weedy var. frutescens, cultivated var. crispa, and weedy var. crispa were 0.415, 0.606, 0.308, and 0.480, respectively. Both weedy accessions exhibited higher GD and PIC values than their cultivated types in East Asia. The new SSR primers of Perilla species reported in this study may provide potential genetic markers for population genetics to enhance our understanding of the genetic diversity, genetic relationship and population structure of the cultivated and weedy types of P. frutescens in East Asia. In addition, new Perilla SSR primers developed from RNA-seq can be used in the future for cultivar identification, conservation of Perilla germplasm resources, genome mapping and tagging of important genes/QTLs for Perilla breeding programs.

• Journal of Microbiology and Biotechnology
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• v.34 no.5
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• pp.1146-1153
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• 2024

The increasing economic losses associated with growth retardation caused by Enterocytozoon hepatopenaei (EHP), a microsporidian parasite infecting penaeid shrimp, require effective monitoring. The internal transcribed spacer (ITS)-1 region, the non-coding region of ribosomal clusters between 18S and 5.8S rRNA genes, is widely used in phylogenetic studies due to its high variability. In this study, the ITS-1 region sequence (~600-bp) of EHP was first identified, and primers for a polymerase chain reaction (PCR) assay targeting that sequence were designed. A newly developed nested-PCR method successfully detected the EHP in various shrimp (Penaeus vannamei and P. monodon) and related samples, including water and feces collected from Indonesia, Thailand, South Korea, India, and Malaysia. The primers did not cross-react with other hosts and pathogens, and this PCR assay is more sensitive than existing PCR detection methods targeting the small subunit ribosomal RNA (SSU rRNA) and spore wall protein (SWP) genes. Phylogenetic analysis based on the ITS-1 sequences indicated that the Indonesian strain was distinct (86.2% nucleotide sequence identity) from other strains collected from Thailand and South Korea, and also showed the internal diversity among Thailand (N = 7, divided into four branches) and South Korean (N = 5, divided into two branches) samples. The results revealed the ability of the ITS-1 region to determine the genetic diversity of EHP from different geographical origins.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.15 no.1
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• pp.29-37
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• 2012

Purpose: To examine the prevalence of $Clostridium$ $difficile$ ($C.$ $difficile$) colonization (CDC) and potential neonatal determinants of CDC in hospitalized preterm infants. Methods: Fecal samples were serially collected within 72 h after birth and at 1, 2, and 4-6 weeks of age from preterm infants in the neonatal intensive care units (NICUs) of two different university hospitals. Total bacterial DNA was extracted from each fecal sample from 49 infants, and polymerase chain reaction (PCR) was performed with primers for the 16S gene of $C.$ $difficile$ and the toxin A and toxin B genes. The correlation between the results of $C.$ $difficile$ PCR assays and the clinical characteristics of the infants was analyzed. Results: The prevalence rates of CDC were 34.7, 37.2, 41.3, and 53.1% within 72 h after birth and at 1, 2, and 4.6 weeks of age, respectively. The toxin positivity rate was significantly higher in the infants with persistent CDC than in those with transient CDC (8/12 [66.7%] vs. 6/25 [24.5%] ($p$=0.001). Among the various neonatal factors, only the feeding method during the first week after birth was significantly associated with persistent CDC. Exclusive breast-milk feeding (EBMF) significantly decreased the risk of persistent CDC compared to formula or mixed feeding (adjusted odds ratio: 0.133, 95% confidence interval: 0.02-0.898, $p$=0.038). Conclusion: The prevalence of CDC increased with the duration of hospitalization in preterm infants in the NICU. EBMF during the first week after birth in hospitalized preterm infants may protect against persistent CDC.