• Reproductive and Developmental Biology
• /
• v.28 no.1
• /
• pp.37-43
• /
• 2004

This study was carried out to investigate PSS (Porcine Stress Syndrome) with the PSE (Pale, Soft, Exudative) in 319 different pigs(Yorkshire 150; Landrace 89 and Duroc 80). The PCR-RFLP method was adapted to detect the ryanodine receptor (RYR 1) gene mutation and to estimate the genotype frequency of the RYR1 gene in breeding pig population. The DNA samples were collected from hair follicles of pigs of Yorkshire, Landrace and Duroc. After DNA amplification by PCR, the PCR products were digested by restriction enzyme, Cfo I. Primary PCR products of ryanodine receptor gene were length of 659 bp in hair follicle and their second PCR products were length of 522 bp in hair follicle. The exon region (522 bp) including point mutation ($C \arrow T; Arg \arrow Cys$) in the porcine ryanodine receptor gene, which is a causal mutation for PSS, was digested with Cfo I restriction enzyme. The RYR1 gene was classifed into three genotypes by agarose gel electrophoresis. The normal homozygous (NN) individuals showed two DNA fragments consisted of 439 and 83 bp. The mutant homozygous (nn) individuals showed only one DNA fragment 522 bp. In addition, all three fragments (522, 439 and 83 bp) were showed in heterozygous (Nn) carrier animals. The normal homozygous (NN), heterozygous (Nn) and mutant homozygous (nn) were 98.00, 2.00 and 0.00% in Yorkshire pigs, 87.64, 11.24 and 1.12% in Landrace, 100.00, 0.00 and 0.00% in Duroc, respectively. The gene frequencies of N and n were 0.990 and 0.010 in Yorkshire pigs, 0.933 and 0.067 in Landrace, 1.000 and 0.000 in Duroc, respectively.

• Asian Pacific Journal of Cancer Prevention
• /
• v.16 no.5
• /
• pp.1987-1992
• /
• 2015

The aim of this research was to investigate the possible association between gastric carcinoma (GC) and polymorphisms of the IL-$1{\beta}$ gene in the Kashmiri population using peripheral blood DNA from 150 gastric carcinoma cases and 250 population controls with detailed data for clinicopathological characteristics of the disease. Two SNPs in the IL-$1{\beta}$ gene were selected for this study. Expression of IL-$1{\beta}$ was studied in 50 gastric carcinoma cases using immunohistochemistry and RT-PCR and then correlated with genotype. The frequency of the IL-$1{\beta}$-511 C allele was significantly higher in the GC case group (53.3%) than in controls (45.4%) with an odds ratio (OR) of 0.73 and a P value of 0.03. Multivariate regression analysis showed associations of gastric carcinoma with mutant form of IL-$1{\beta}$-511 TT (OR 0.309; P value <0.001) and the CC genotype of IL-$1{\beta}$-31 (OR 0.313; P value of 0.002). Haplotype analysis of IL-$1{\beta}$-31 and IL-$1{\beta}$-511 showed decreased association of IL-$1{\beta}$-31 T with IL-$1{\beta}$-511 C with gastric carcinoma (OR 0.728; P value 0.03). Expression study of 50 samples by immunohistochemistry (IHC) and RT-PCR showed association with grade III and stage III+IV. After correlating the expression with polymorphism no association was found.

• Korean Journal of Microbiology
• /
• v.28 no.1
• /
• pp.47-54
• /
• 1990

In order to investigate physiological characteristics of fusants by interspecific protoplast fusion of the genus Cellulomonas, protoplasts of Cellulomonas flavigena NCIB 12901 and Cellulomonas bibula NCIB 8142 were fused and cell wall regenerated. To give gene maker, C. bibula was treated with 500 ug/ml NTG for 1 hr and arginine requiring auxotrophic mutants were isolated. Protoplasts of the genus Cellulomonas were obtained by treatment with $600{\mu}{\textrm{g}}$/ml lysozyme, and 0.5M sorbitol was optimal for osmotic stabilizer on protoplast fromation. Protoplast fusion was enhanced by 40% PEG)M.W.6,000) containing 25 mM $CaCl_{2}$ at $30^{\circ}C$ for 30 min and fusion frequency between C. bibula and C. flavigena was $5\times 10^{-4}$. Processes of protoplast formation, cell wall regeneration and protoplast fusion were obsdrved by scanning electron microscope. By comparing enzyme activities of cellulase, exocellobiohydrolase, .betha.-glucosidase of the parent strains of Cellulomonas with those of thier mutants and fusants, fusants with increased enzyme activity were obtained. By the studies on nutritional requirement, antibiotic resistance, cellulolytic enzyme activities, type of peptidoglycan and motility of two mutants and fusants, fusants were proved to be recombinant of both mutant strains.

• Asian Pacific Journal of Cancer Prevention
• /
• v.16 no.11
• /
• pp.4583-4587
• /
• 2015

Background: The objectives of this study aimed to detect a CYP2B6 polymorphism in de novo cases of acute myeloid leukemia patients and identify any role in disease progression and outcome. Materials and Methods: DNA was isolated from peripheral blood of 82 newly diagnosed acute myeloid leukemia cases and the CYP2B6 G15631T gene polymorphism was assayed by PCR restriction fragment length polymorphism (PCR-RFLP). Results: The frequency of the GG genotype (wild type) was 48 (58.5%) and that of the mutant type T allele was 34 (41.9%). GT genotype heterozygous variants were found in 28 (34%), and TT genotype homozygous variants in 6 (7.3%) cases. We found no significant association between the CYP2B6 G15631T polymorphism and complete response (CR) (p-value=0.768), FAB classification (p-value=0.51), cytogenetic analysis (p-value=0.673), and overall survival (p-value=0.325). Also, there were no significant links with early toxic death (p-value=0.92) or progression-free survival (PFS) (p-value=0.245). Conclusions: Our results suggest that the CYP2B6 polymorphism has no role in disease progression, therapeutic outcome, patient free survival, early toxic death and overall survival in acute myeloid leukemia patients.

• BMB Reports
• /
• v.51 no.10
• /
• pp.481-483
• /
• 2018

Glioblastoma (GBM) is the most common and aggressive form of human adult brain malignancy. The identification of the cell of origin harboring cancer-driver mutations is the fundamental issue for understanding the nature of GBM and developing the effective therapeutic target. It has been a long-term hypothesis that neural stem cells in the subventricular zone (SVZ) might be the origin-of-cells in human glioblastoma since they are known to have life-long proliferative activity and acquire somatic mutations. However, the cell of origin for GBM remains controversial due to lack of direct evidence thereof in human GBM. Our recent study using various sequencing techniques in triple matched samples such as tumor-free SVZ, tumor, and normal tissues from human patients identified the clonal relationship of driver mutations between GBM and tumor-free SVZ harboring neural stem cells (NSCs). Tumor-free SVZ tissue away from the tumor contained low-level GBM driver mutations (as low as 1% allelic frequency) that were found in the dominant clones in its matching tumors. Moreover, via single-cell sequencing and microdissection, it was discovered that astrocyte-like NSCs accumulating driver mutations evolved into GBM with clonal expansion. Furthermore, mutagenesis of cancer-driving genes of NSCs in mice leads to migration of mutant cells from SVZ to distant brain and development of high-grade glioma through the aberrant growth of oligodendrocyte precursor lineage. Altogether, the present study provides the first direct evidence that NSCs in human SVZ is the cell of origin that develops the driver mutations of GBM.

• Korean Journal of Food Science and Technology
• /
• v.25 no.4
• /
• pp.366-372
• /
• 1993

This study mainly designed to high quality of mirin production by using protopast fusion. In order to enhance the acid carboxypeptidase (ACPase) activity by the method of protoplast fusion. In order to enhance the acid carboxypeptidase (ACPase) activity by the method of protopalst fusion, the mutants, Aspergillus oryzae 9-12 and Aspergillus shirosamii IFO 6082-60 were selected by mutation among various mutants. Protoplast of Aspergillus oryzae 9-12 and Aspergillus shirousamii IFO 6082-60 were formed effectively by incubation of the mixtures of chitinase (10mg/ml), cellulase (10mg/ml) and zymolase 20T (5mg/ml). For protopalst fusion, the mixture of two mutant were fused to effective under the optimum conditions by solutions containing 30% PEG 6,000, 0.01M $CaCl_2\;2H_2O$, 0.6M KCl and 0.05M glycine. Fusion frequency was 0.71% and fusant, F-50 appeared ACPase activity of 20,800 unit/g which has 1.5 times higher than that of each mutants.

• Journal of Microbiology and Biotechnology
• /
• v.3 no.4
• /
• pp.232-237
• /
• 1993

A strain of yeast which can convert starch directly to ethanol was developed by the intergeneric protoplast fusion between Schwanniomyces alluvius possessing $\alpha$ amylase as well as glucoamylase with debranching activity and FSC-14-75 which previously had been formed from a heterologous transformation and subsequent intergeneric protoplast fusion. Fusants were selected on minimal medium after protoplasts of auxotrophic mutant of S. alluvius fused with heat-treated protoplasts of FSC-14-75 in the presence of 30%(w/v) PEG and 20 mM $CaCl_2$. The fusion frequency was in the range of $10^{-6}$ order. All fusants tested were intermediate types of parental strains for carbon compound assimilation, and their cell volumes were approximately 1.1 times larger than FSC-14-75 and 1.8 times larger than S. alluvius. The fusants were unable to sporulate like FSC-14-75, while S. alluvius could sporulate. In flask scale the most promising fusant, FSCSa-R10-6, produced 7.83%(v/v) and 10.17%(v/v) ethanol from 15% and 20% of liquefied potato starch, respectively, indicating that the fermetation efficiency of each case increased 1.2 times and 1.6 times than that of FSC-14-75. The elution pattern on DEAE-cellulose chromatography showed that FSCSa-R10-6 has four distinct amylase peaks of which two peaks originated from S. alluvius and the other two from FSC-14-75. These results suggest that the enhanced fermentation efficiency of the fusant might be due to almost-complemented parental amylases.

• The Korean Journal of Physiology and Pharmacology
• /
• v.7 no.3
• /
• pp.157-162
• /
• 2003

Our earlier studies found a significant correlation between the activities of ranitidine N-oxidation catalyzed by hepatic flavin-containing monooxygenase (FMO) and the presence of mutations in exon 4 (E158K) and exon 7 (E308G) of the FMO3 gene in Korean volunteers. However, caffeine N-1 demethylation (which is also partially catalyzed by FMO) was not significantly correlated with these FMO3 mutations. In this study, we examined another common mutation (V257M) in exon 6 of FMO3 gene. The V257M variant, which is caused by a point mutation (G769A), was commonly observed (13.21% allele frequency) in our subjects (n=159). This point mutation causes a substitution of $Val^{257}$ to $Met^{257}$, with transformation of the secondary structure. The presence of this mutant allele correlated significantly with a reduction in caffeine N-1-demethylating activity, but was not correlated with the activity of N-oxidation of ranitidine. In a family study, the low FMO activity observed in a person heterozygous for a nonsense mutation in exon 4 (G148X) and heterozygous for missense mutation in exon 6 (V257M) of FMO3 was attributed to the mutations. Our results suggest that various point mutations in the coding regions of FMO3 may influence FMO3 activity according to the probe substrates of varying chemical structure that correlate with each mutation on the FMO3 gene.

• Journal of Periodontal and Implant Science
• /
• v.42 no.5
• /
• pp.151-157
• /
• 2012

Purpose: Cyclooxygenase (COX) enzyme catalyzes the production of prostaglandins, which are important mediators of tissue destruction in periodontitis. Single nucleotide polymorphisms of $COX_2$ enzyme have been associated with increasing susceptibility to inflammatory diseases. The present study evaluates the association of two single nucleotide polymorphisms in $COX_2$ gene (-1195G>A and $8_{473}$C>T) with chronic periodontitis in North Indians. Methods: Both SNPs and their haplotypes were used to explore the associations between $COX_2$ polymorphisms and chronic periodontitis in 56 patients and 60 controls. Genotyping was done by polymerase chain reaction followed by restriction fragment length polymorphism. Chi-square test and logistic regression analysis were performed for association analysis. Results: By the individual genotype analysis, mutant genotypes (GA and AA) of $COX_2$-1195 showed more than a two fold risk (odds ratio [OR]>2) and $COX_2$ $8_{473}$ (TC and CC) showed a reduced risk for the disease, but the findings were not statistically significant. Haplotype analysis showed that the frequency of the haplotype AT was higher in the case group and a significant association was found for haplotype AT (OR, 1.79; 95% confidence interval, 1.03 to 3.11; P=0.0370) indicating an association between the AT haplotype of $COX_2$ gene SNPs and chronic periodontitis. Conclusions: Individual genotypes of both the SNPs were not associated while haplotype AT was found to be associated with chronic periodontitis in North Indians.

• Journal of Life Science
• /
• v.20 no.3
• /
• pp.453-456
• /
• 2010

Leukemia is the abnormal increase of hematopoietic progenitor cells in tissues, resulting in anemia, increased susceptibility to infection and impaired blood clotting. The adenosine deaminase (ADA) gene is an important druggable target for the treatment of leukemia patients. Genetic and molecular analyses were performed to determine the effects of ADA gene mutations in 20 leukemia patients in the Korean population. To analyze the relationship between genotype and phenotype, the ADA genomic DNAs - including 1,092 bp of 12 exons and partial intron sequences flanking each exon - were sequenced and compared. In this study, the known mutations in other diseases, more than 50 mutations already reported in patients with severe combined immunodeficiency disease (SCID) and autism, were not found, but two novel mutations in leukemia patients were discovered. They include one nonsense mutation (A>C at nt position 478, F101F) and one missense mutation (G>A at nt position 778, E260K). One missense mutation (G>A at nt position 22, D8Y) was also detected in 20 normal control patients (allelic frequency of 7.5%). Interestingly, subjects in the Korean population retained 2 bp insertion at the intron 6 (IVS6-52insGC), something that has never been shown in other populations. The genetic study to find out the correlation between the mutant alleles and leukemia patients revealed no association statistically (p>0.05). The mutation found in leukemia needs further study to determine its possibility as a molecular marker for the diagnosis of leukemia.