• Mycobiology
• /
• v.48 no.5
• /
• pp.427-430
• /
• 2020

Ganoderma lingzhi is a well-known source of natural fungal medicines which has been given for the treatment of several diseases. China is one of the major commercial producers of Ganoderma mushroom worldwide. However, with the expansion of the commercial cultivation, the occurrence of the fungal diseases on G. lingzhi has also been increased. The green mold disease symptoms were observed in the cultivation base of G. lingzhi in Zuojia Town, Jilin City, Jilin Province, China, causing the basidiomes to be rotten and withered, and the green mycelium layer generated gradually. The pathogenicity tests showed the same symptoms as appeared naturally in Zuojia mushroom base. Morphology characters revealed conidia green, ellipsoid, globose, 2.56-4.83 × 2.09-4.22 ㎛, length-width ratio was 1.1-1.2 (n = 10). Conidiophores trichoderma-like, often asymmetry, branches solitary, paired or in whorls of 3 phialides formed solitary, paired or in whorl, variable in shape, lageniform, sometimes ampulliform or subulate. While using molecular methodology, comparing with the sequences of Trichoderma hengshanicum from GenBank, the analyzed sequence showed 97.32% homology with the RPB2 sequences, 100% with the TEF1-a sequences. A fungus isolated from the diseased tissues was identified based on morphology and molecular studies as T. hengshanicum. This is the first report of T. hengshanicum causing the green mold disease of G. lingzhi in China.

• Korean Journal of Agricultural Science
• /
• v.48 no.2
• /
• pp.353-365
• /
• 2021

Previously, we reported that tissue factor (Tf) was included in the list of differentially expressed genes as an upregulated gene in a rejected porcine heart after xenotransplantation into monkey. In this study, we analyzed that expression of Tf in aortic endothelial cells (pAEC) isolated from alpha 1,3-galactosyltransferase knockout pig in response to allogeneic porcine serum and xenogeneic human serum. The consequence was significant upregulation of Tf expression by responding to human serum compared with porcine serum. To analyze the function of Tf gene as a promoter, we constructed reporter vectors for expression of luciferase linked to 1,246 and 787 base pairs of porcine Tf (pTF1246 and pTF787), and 535 base pairs of human TF (hTF535) sequences including putative promoter regions and AP-1 biding site at the 5' end. The reporter vectors were transfected into pAEC including cytomegalovirus enhancer/chicken β-actin (CAG)-luciferase vector as a control. Luciferase assay showed that all of the promoters were insufficient to express luciferase compared with CAG promoter in basic culture conditions. Notably, pTF1246, pTF787, and hTF535 led to a significant increase of luciferase expression in response to human serum compared with porcine serum while no change of CAG. pTF1246 and pTF787 showed higher expression than hTF535. Taken together, our findings suggest that pTF1246 and pTF787 promoters could mediate target gene expression specifically at xenogeneic stress condition.

• Biomedical Science Letters
• /
• v.14 no.1
• /
• pp.47-53
• /
• 2008

Hox genes are known to be transcription factors controlling vertebrate pattern formation along the anteroposterior body axis by regulating many target gene expressions during vertebrate embryogenesis. In order to isolate in vivo Hox responsive target genes, ChIP-cloning technique has been applied using Hoxc8 antibody. Here murine embryo of day 11.5 post coitum (E11.5) highly expressing Hoxc8 gene was used after removing head and tail portions where Hoxc8 is rarely expressing. After fixation with formaldehyde, the chromatin DNAs harboring bound proteins were isolated. After sonication, about 0.5- to 1 Kb chromatin DNAs were immunoprecipitated with anti Hoxc8 antibody. After removing the bound proteins with proteinase K, DNAs were isolated, cloned into the pBluescsript II SK vector, and then sequenced. Total 33 random clones sequenced were anlalyzed to be located at 12 different genomic regions. Among these, 8 turned out to be introns and 4 were intergenic regions localized in random chromosomes. The base composition of total cloned genomic sequences (6608 bp) were AT-rich, i.e., 40% GC. When the Hoxc8 core binding sites, such as TAAT, ATTA, TTAT, and ATAA were analyzed total number of 55, 45, 54, and 55 were found, respectively, which are than twice as many as expected number of 26. Although this in silico analysis does not mean that the ChIP-cloned sequence is real Hoxc8 regulatory element in vivo, these results strongly imply that the DNA fragments cloned through chromatin immunoprecipitation could be very much likely the putative Hoxc8 downstream target genes.

• Journal of Trauma and Injury
• /
• v.31 no.1
• /
• pp.1-5
• /
• 2018

Purpose: Although there has been substantial progress for the treatment of thoracic trauma, the mortality of the penetrating neck injury is still high, has been reported about 10-15%. However, there has not been a report which is reflecting Korean medical present. We retrospectively analyzed the penetrating neck injury patients based on the Korean Trauma Data Base. Methods: Between December 2013 and June 2017 at the trauma center of the Pusan National University Hospital, Busan, Korea, total of 36 patients with isolated low-velocity penetrating neck injuries were included. We analyzed the patients' age, gender, injury mechanism and causes by medical chart review. Results: Among total of 36 patients, 26 (72.2%) were male and 10 (27.8%) were female. Homicidal neck injuries were most common, followed by accidental and suicidal injuries (47% vs. 33% vs. 19%, respectively). All penetrating injuries in our study were low-velocity trauma such as following: knife (n=16, 44.4%); glass or glass bottle (n=11, 30.6%); scissors (n=4, 11.1%); grinder (n=2, 5.6%); and three (8.3%) of miscellaneous injuries. Twenty-seven (75.0%) patients underwent emergency surgery, and only one (2.8%) patient underwent elective surgery. Eleven (30.6) patients were diagnosed with superficial injuries, including six patients who had conservative treatment. Twelve (33.3%) patients had arterial injuries and 10 (27.8%) patients had venous injuries. The patients who had deep injuries showed significant difference against the patient with superficial injury (98.0 vs. 129.1, p=0.008). Conclusions: Low velocity penetrating injury confined to the neck is able to be successfully treated with prompt surgical management. Regardless of the conditions which are evaluated at emergency department, all penetrating neck injury patients should be regarded as urgent surgical candidates.

• Clinical and Experimental Pediatrics
• /
• v.58 no.1
• /
• pp.20-27
• /
• 2015

Purpose: We investigated the molecular types of uropathogenic Escherichia coli (UPEC) by using conventional phylogrouping, multilocus sequence typing (MLST), and fimH genotyping. Methods: Samples of patients younger than 18 years of age were collected from the Chung-Ang University Hospital over 2 years. Conventional phylogenetic grouping for UPEC strains was performed by polymerase chain reaction (PCR). Bacterial strain sequence types (STs) were classified on the basis of the results of partial sequencing of seven housekeeping genes. In addition, we analyzed nucleotide variations in a 424-base pair fragment of fimH, a major virulence factor in UPEC. Results: Sixty-four UPEC isolates were analyzed in this study. Phylogenetic grouping revealed that group B2 was the most common type (n=54, 84%). We identified 16 distinctive STs using MLST. The most common STs were ST95 (35.9%), ST73 (15.6%), ST131 (12.5%), ST69 (7.8%), and ST14 (6.3%). Fourteen fimH allele types were identified, of which 11 had been previously reported, and the remaining three were identified in this study. f1 (n=28, 45.2%) was found to be the most common allele type, followed by f6 and f9 (n=7, 11.3% each). Comparative analysis of the results from the three different molecular typing techniques revealed that both MLST and fimH typing generated more discriminatory UPEC types than did PCR-based phylogrouping. Conclusion: We characterized UPEC molecular types isolated from Korean children by MLST and fimH genotyping. fimH genotyping might serve as a useful molecular test for large epidemiologic studies of UPEC isolates.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.56 no.4
• /
• pp.349-360
• /
• 2011

The objectives of present study were to characterize the peptides which were isolated from Korean fermented soybean paste, chungkukjang, and to determine their antioxidant activities. Four fractions were collected from the methanol extract of chungkukjang by using a recycling preparative HPLC. Among fractions, Fr-2 was identified to be highly potent free radical scavenging activity in the assay of 1,1-diphenyl-2-picryl-hydrazyl (DPPH) and nitroblue tetrazolium(NBT)-reduction inhibition. Base on antioxidant effects, fraction Fr-2 was employed for the refraction with a prep-column and separated into five fractions of which two fractions were identified to have higher antioxidant activity. To confirm the amino acid constituents of antioxidant fractions Fr-2-2 and Fr-2-3 were analyzed, and eight kinds of amino acids such as aspartic acid, threonine, serine, glutamic acid, glycine, lysine, histidine, and arginine were identified as the constituent amino acids. Antioxidant activities of the separated peptides were further assessed cell viability with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl terazolium bromide (MTT), and fluorescence-activated cell sorting (FACS) analysis of H4IIE cells treated with hydrogen peroxide (H2O2). Chungkukjang peptides have shown their ability to protect H4IIE rat hepatoma cells against H2O2- induced oxidative stress by concentration and time-dependent manner. Therefore, These results indicated that fermented soybean paste chungkukjang will be promoted the antioxidant and radical scavenging activities, and beneficial for health. The antioxidant peptide fractions Fr-2-2 and Fr-2-3 were denominated as P-NICS-1 and P-NICS-2, respectively. However, further studies were required to clarify their amino acid sequences and molecular properties, and physiological significances.

• Applied Biological Chemistry
• /
• v.44 no.3
• /
• pp.153-161
• /
• 2001

In order to isolate genes involved in ecdysteroids biosynthesis in plants, total RNA was isolated from Achyranthes japonica Nakai, and RT-PCR was performed using degenerate primers selected based on the results of multi-alignment of four cytochrome P450 genes from plants and a putative ecdysone 20-hydroxylase gene from an insect. Fourteen partial cDNA clones showing unique base sequences were obtained, out of which six showed homologies at the levels of nucleotide and amino acid sequences to the other cytochrome P450 genes known to be involved in the ecdysteroid biosynthesis. Of the six clones, four showed relatively high homologies to a putative ecdysone 20-hydroxylase gene isolated from an insect.

• Journal of Microbiology and Biotechnology
• /
• v.25 no.9
• /
• pp.1449-1459
• /
• 2015

A novel proteolytic enzyme with fibrinolytic activity, FSP3, was purified from the recently isolated Streptomyces sp. P3, which is a novel bacterial strain isolated from soil. FSP3 was purified to electrophoretic homogeneity by ammonium sulfate precipitation, anion exchange, and gel filtration. FSP3 is considered to be a single peptide chain with a molecular mass of 44 kDa. The maximum activity of the enzyme was observed at 50℃ and pH 6.5, and the enzyme was stable between pH 6 and 8 and below 40℃. In a fibrin plate assay, FSP3 showed more potent fibrinolytic activity than urokinase, which is a clinical thrombolytic agent acting as a plasminogen activitor. The activity was strongly inhibited by the serine protease inhibitor PMSF, indicating that it is a serine protease. Additionally, metal ions showed different effects on the activity. It was significantly suppressed by Mg²⁺ and Ca²⁺ and completely inhibited by Cu²⁺, but slightly enhanced by Fe²⁺. According to LC-MS/MS results, its partial amino acid sequences are significantly dissimilar from those of previously reported fibrinolytic enzymes. The sequence of a DNA fragment encoding FSP3 contained an open reading frame of 1287 base pairs encoding 428 amino acids. FSP3 is a bifunctional enzyme in nature. It hydrolyzes the fibrin directly and activates plasminogen, which may reduce the occurrence of side effects. These results suggest that FSP3 is a novel serine protease with potential applications in thrombolytic therapy.

• Biomedical Science Letters
• /
• v.4 no.2
• /
• pp.137-142
• /
• 1998

We examined mRNAs, isolated from the rat brain, to ascertain if there is any polymorphism for S-100 beta protein gene. As templates for polymerase chain reaction (PCR) the reverse-transcribed cDNA from the rat brain or phage DNAs isolated from the rat brain cDNA libraries were used. Although PCR products turned out to be exactly same as the expected size based on the previously reported mRNA sequence a single base substitution (CAT to CAC) was identified at nucleotide level. This change was considered as polymorphism since it did not cause any change of the primary structure for S-100 beta protein. This result should facilitate the understanding of the overall structure of the gene for S-100 beta protein.

• Research in Plant Disease
• /
• v.22 no.1
• /
• pp.55-58
• /
• 2016

A Cucumber mosaic virus (named CMV-Tr1) isolated from the white clover (Trifolium repens) showing mosaic and malformation that found in a pepper field. Cucumber mosaic virus was identified through confirmation with PT-PCR, PCR-restriction fragment length polymorphism, and sequence analysis of coat protein (CP) gene. CMV-Tr1 mosaic symptom on the upper leaves of five tobacco species including Nicotiana benthamiana, Cucumis sativus, Physalis angulata, and Solanum lycopersicon. In Chenopodium quinoa and Vigna unguiculata the isolate showed local lesions in inoculated leaves. CMV-Tr1 compared with CMV-As in the sequence identity of CP gene. CMV-Tr1 showed 98.9% and 99.5% homologies at nucleotide and amino acid levels, respectively. Phylogenetic analysis of the CP gene indicated that CMV-Tr1 belongs to the CMV subgroup IB base on the CP. To our knowledge, this is the first report of CMV in T. repens in Korea.