• Journal of Sericultural and Entomological Science
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• v.53 no.1
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• pp.44-49
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• 2015

We characterized tissue specific-expressed genes in the posterior silk gland of Bombyx mori using by the Annealing Control Primer based differential display-PCR manner. In this study, we isolated 34 differentially expressed PCR amplicons, which one of these was identified as a novel transcript named as ACP-16 (366 bp), its expression was observed only in the posterior silk glands by Northern blot analysis. To determine promoter region of the ACP-16, we isolated and analyzed a phage DNA having 1.7 kb-long genome DNA including the open reading flame and 5'- upstream untranslated region of the ACP-16 gene from a genomic DNA library. We have estimated a promoter region of the ACP-16 gene by a web promoter prediction engine, which locates -750 ~ -165 from translation initiation site (ATG, +1). ACP-16 gene is necessary to more studies about critical biological role in order to apply the silkworm's transgenic system.

• Journal of Sericultural and Entomological Science
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• v.52 no.1
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• pp.39-44
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• 2014

We isolated highly-expressed genes in the posterior silk glands of silkworm on a previously study, which one of these was identified as RNA binding protein-1 homologue (RBP-1) gene. In this study, we investigated gene expressional characteristics of the RBP-1 depending on silkworm development stages and several tissues of the larvae, respectively. Northern blot hybridization analysis showed that the RBP-1 gene was expressed high in larval and pupal periods, and highly expressed than endogenous internal control gene (BmA3) on all tested larval tissues. In addition, we isolated and analyzed a phage DNA having 1,660 bp-long promoter region of the RBP-1 gene from a genomic DNA library. To study the RBP-1 gene promoter activity, RBP-1 (-740/+ 30) was amplified by PCR and subcloned into a pGL3 basic vector to generate pGL-RBP1. A luciferase report vector carrying RBP-1 gene promoter (770 bp) was tested by luciferase assay in Sf9 cells. In the result, the RBP-1 gene promoter was more efficient than constitutive promoter (BmA3) by approximately ten percent.

• Journal of Microbiology and Biotechnology
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• v.13 no.6
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• pp.1013-1017
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• 2003

A thermophilic nonspore-forming rod isolated from hay compost in Korea was subjected to a taxonomic study. The microorganism, designated as $SC-1^T$, was identified as a nitrate-reducing and nonmotile bacterium. Although the strain was negatively Gram-stained, a KOH test showed that the strain $SC-1^T$ belonged to a Gram-positive species. Growth was observed between 45 and $70^{\circ}C$. The optimal growth temperature and pH were $60^{\circ}C$ and pH 7.5, respectively. The G+C content of the genomic DNA was 65 mol% and the major quinone types were MK-6 and MK-7. A phylogenetic analysis based on 16S rDNA sequences revealed that the strain $SC-1^T$ was most closely related to Symbiobacterium thermophilum. However, the level of DNA-DNA relatedness between strain $SC-1^T$ and the type strain for Symbiobacterium thermophilum was approximately 30%. Accordingly, on the basis of the phenotypic traits and molecular systematic data, the strain $SC-1^T$ would appear to represent a new species within the genus Symbiobacterium. The type strain for the new species is named $SC-1^T$ ($=KCTC\;0307BP^T;\;DSM15906^T$).

• Applied Biological Chemistry
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• v.45 no.4
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• pp.185-189
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• 2002

Along with the worldwide rapid increase of the cultivation area and commercial production of genetically modified (GM) crops, the amount of GM grains imported to Korea has also been increasing. Roundup-Ready soybean (RRS) was introduced with 5-enolpyruvyl shikimate-3-photphate synthase (EPSPS) gene derived from Agrobacterium CP4 to confer the resistance to herbicide, glyphosate. In this study, we tried to develop PCR-based analytical method to detection the presence of RRS among non-GM soybeans. In order to detect RRS specifically, oligonucleotide primers were specifically designed based on the nucleotide sequence of EPSPS transgene. Qualitative PCR method was established and its specificity and accuracy were confirmed by analysing the nucleotide sequence of PCR DNA fragments. Bioassay was also conducted by spraying glyphosate at seedling stage. Survived individuals showed obvious resistance to Roundup Ready, however all of non-GM seedlings died in two weeks after spray. Conclusively, the highly selective detection systems for RRS were successfully established by both PCR using specific primers to EPSPS transgene and bioassay using the herbicide resistance of RRS. In addition to, the imported soybean showed to be mixed to several varieties regarding to 100-seed weight and hilum color.

• Journal of Embryo Transfer
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• v.21 no.4
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• pp.287-297
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• 2006

Concentration of hormones and blood components at the last fatting stage was changed before slaughter in Hanwoo steers and bulls. Two months before slaughter and shipment, concentration of cortisol and creatinine was increased, but that of calcium was decreased. Concentration of insulin growth factor-1 (IGF-1) was decreased after shipment, and inorganic phosphorus (IP) was decreased at slaughter. It is unclear that changes of concentration in between 2 months before slaughter and shipment were either caused by aging or stresses (abstinence, environmental change, blood drawing, and shipment). Changes of blood concentration between shipment and slaughter may be accounted for overall responses from abstinence, shipment, and unfamiliar environment. A positive correlation between 2 months before slaughter and before shipment was detected for IGF-1, total protein (TP), albumin, creatinine, high density lipoprotein cholesterol (HDLC), and globulin in steers, and creatinine and globulin in bulls. A positive correlation between 2 month before slaughter and slaughter was detected for IGF-1, blood urea nitrogen (BUN), IP and HDLC in steers, and creatinine in bulls. A positive correlation between before shipment and slaughter was detected for testosterone, IGF-1, creatinine, triglyceride, HDLC and globulin in steers, and TP, creatinine, HDLC and globulin in bulls.

• Clinical and Experimental Pediatrics
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• v.48 no.8
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• pp.877-880
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• 2005

Purpose : Glycogen storage disease type Ia(GSD Ia) is an autosomal recessive disorder caused by the deficiency of glucose-6-phosphatase(G6Pase). The aim of the study was to investigate the spectrum of G6Pase gene mutations and relationship between genotype and clinical findings in Korean patients with GSD Ia. Methods : Genomic DNA was extracted from peripheral leukocytes of 20 patients with GSD Ia. The five exons of G6Pase gene were amplified and PCR products were directly sequenced. The frequency of short stature, hypoglycemia, hypercholesterolemia, hyperuricemia, hypercalciuria, nephrocalcinosis and hepatic adenoma was compared between 727G>T homozygotes and 727G>T compound heterozygotes. Results : A total of 5 different mutations were identified. The most common mutation was the 727G>T with an allele frequency of 80%. All patients were either homozygous(12/20) or heterozygous(8/20) for the 727G>T mutation. G122D was found in 3 patients, P178A in 1, G222R in 2, and S339R in 2. There was no difference in the frequency of short stature, hypoglycemia, hypercholesterolemia, hyperuricemia, nephrocalcinosis, and hepatic adenoma between 727G>T homozygotes and heterozygotes. Conclusion : Diagnosis of GSD Ia can be based on clinical and biochemical abnormalities combined with mutation analysis instead of enzymatic diagnosis that requires liver biopsy. Homozygosity for the 727G>T does not seem to alter the disease phenotype as compared with the heterozygous state.

• Microbiology and Biotechnology Letters
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• v.35 no.1
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• pp.1-10
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• 2007

The 44 endophytic bacterial strains were isolated from surface-sterilized root of rice cultivated in seven different locations of Chungcheong province, Korea. Each isolate was introduced into rice seedlings grown gnotobiotically by inoculating scissor-cut first true leaf with cell suspensions, and the colonization capacity of each isolate in root tissue was analyzed at 7 days after inoculation. Sixteen out of 44 isolates were re-isolated from root successfully with the frequency of $10^{3-5}$ CFU/g tissue. Interestingly, seven out of 16 isolates were identified as Burkholderia species. The identity between inoculated strains and re-isolates was confirmed by genomic finger-printing and 16S rDNA sequence analysis. By a confocal laser scanning microscopic observation it was revealed that KJ001 strain, one of the sixteen isolates tagged with gfp colonized in root tissue especially around xylem. Six out of seven Burkholderia strains obtained in this study showed antagonizing activities against seven different fungal pathogens, contain nifH gene, and five of them enhanced growth of cucumber over 30%. The isolates showed no hypersensitive response on tobacco leaves and no pathogenecity in rice. From these results it was found that the endophytic Burkholderia strains will be useful in agriculture to develop a biocontrol agent or a bio-fertilizer.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.1
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• pp.369-374
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• 2014

Background: Colorectal cancer (CRC) is the fourth most common cause of cancer death in the world. Genetic variants in 8q24.21 including rs10505477 and rs6983267 have been hypothesized to be involved in susceptibility to CRC. This study aims to investigate the possible association between these loci and their haplotypes with CRC risk in Iranian population. Materials and Methods: Subjects were recruited from two hospitals in Tehran. The rs10505477 and rs6983267 polymorphisms were genotyped by TaqMan real time PCR using subject genomic DNA, extracted either from formalin-fixed, paraffin-embedded tissue of patients or from blood of the controls by standard methods. Results: A total of 715 subjects (380 CRC patients and 335 matched controls) were genotyped in this study. Allele and genotype analysis of the rs10505477 and rs6983267 polymorphisms by gender, age at diagnosis, tumor location, tumor grade, and tumor node metastasis (TNM) showed no significant association with CRC risk. There was a significant relationship between GG haplotype and susceptibility to age at diagnosis for both <60 and ${\geq}60$ (p=0.0005 and p=0.000004, respectively) and between GT and CRC in the age at diagnosis ${\geq}60$ (Table 3: p=0.031). The GG haplotype was less frequent in CRC patients with the age at diagnosis <60, but was more common in subjects with the age at diagnosis ${\geq}60$. Conclusions: Results of this study suggests that the rs6983267 and rs10505477 polymorphisms alone may not be relevant to CRC risk, but their GG haplotype plays a notable role in age at diagnosis of CRC in the Iranian population.

• Journal of Microbiology and Biotechnology
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• v.26 no.4
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• pp.763-774
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• 2016

Equine gastric ulcer syndrome is one of the most frequently reported diseases in thoroughbred racehorses. Although several risk factors for the development of gastric ulcers have been widely studied, investigation of microbiological factors has been limited. In this study, the presence of Helicobacter spp. and the gastric microbial communities of thoroughbred racehorses having mild to severe gastric ulcers were investigated. Although Helicobacter spp. were not detected using culture and PCR techniques from 52 gastric biopsies and 52 fecal samples, the genomic sequences of H. pylori and H. ganmani were detected using nextgeneration sequencing techniques from 2 out of 10 representative gastric samples. The gastric microbiota of horses was mainly composed of Firmicutes (50.0%), Proteobacteria (18.7%), Bacteroidetes (14.4%), and Actinobacteria (9.7%), but the proportion of each phylum varied among samples. There was no major difference in microbial composition among samples having mild to severe gastric ulcers. Using phylogenetic analysis, three distinct clusters were observed, and one cluster differed from the other two clusters in the frequency of feeding, amount of water consumption, and type of bedding. To the best of our knowledge, this is the first study to investigate the gastric microbiota of thoroughbred racehorses having gastric ulcer and to evaluate the microbial diversity in relation to the severity of gastric ulcer and management factors. This study is important for further exploration of the gastric microbiota in racehorses and is ultimately applicable to improving animal and human health.

• Microbiology and Biotechnology Letters
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• v.40 no.4
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• pp.287-295
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• 2012

Eighty-four endophytic fungal strains were isolated and identified from the roots of halophytes collected in Buan salt marsh. All halophyte samples, such as Suaeda japonica, and Suaeda maritima were isolated from Buan salt marsh. All endophytic fungi isolated were analyzed by sequences of internal transcribed spacer (ITS) containing ITS1, 5.8s and ITS2 region. All endophytic fungi expressed that fungal strains belong to eight orders; Pleosporales (45%), Eurotiales (27%), Incertae sedis (11%), Dothideales (6%), Capnodiales (5%), Hypocreales (5%), and Agaricales (1%). All endophytic fungi were confirmed at the genus level of Ascomycota and Basidiomycota, containing Alternaria, Ascomycota, Aspergillus, Aureobasidium, Cladosporium, Eupenicillium, Fusarium, Gibberella, Hypocrea, Lewia, Macrophoma, Penicillium, Peyronellaea, Phoma, Pleospora, Pleosporales, Pseudeurotium, Schizophyllum, and Talaromyces. Alternaria (21%) and Penicillium (13%) were the dominant endophytic fungal strains. In this study, endophytic fungal strains analyzed from S. japonica and S. maritime, Alternaria (21%), and Penicillium (13%) of Pleosporales and Eurotiales in halophytes were very abundant.