• Molecules and Cells
• /
• v.25 no.2
• /
• pp.265-271
• /
• 2008

Single nucleotide polymorphisms (SNPs) are the most common form of human genetic variation. Non-synonymous SNPs (nsSNPs) change an amino acid. Organic anion transporters (OATs) play an important role in eliminating or reabsorbing endogenous and exogenous organic anionic compounds. Among OATs, hOAT4 mediates high affinity transport of estrone sulfate and dehydroepiandrosterone sulfate. The rapid bone loss that occurs in post-menopausal women is mainly due to a net decrease of estrogen. In the present study we searched for SNPs within the exon regions of hOAT4 in Korean women osteoporosis patients. Fifty healthy subjects and 50 subjects with osteoporosis were screened for genetic polymorphism in the coding region of SLC22A11 (hOAT4) using GC-clamp PCR and denaturing gradient gel electrophoresis (DGGE). We found three SNPs in the hOAT4 gene. Two were in the osteoporosis group (C483A and G832A) and one in the normal group (C847T). One of the SNPs, G832A, is an nsSNP that changes the $278^{th}$ amino acid from glutamic acid to lysine (E278K). Uptake of [$3^H$] estrone sulfate by oocytes injected with the hOAT4 E278K mutant was reduced compared with wild-type hOAT4. Km values for wild type and E278K were $0.7{\mu}M$ and $1.2{\mu}M$, and Vmax values were 1.8 and 0.47 pmol/oocyte/h, respectively. The present study demonstrates that hOAT4 variants can causing inter-individual variation in anionic drug uptake and, therefore, could be used as markers for certain diseases including osteoporosis.

• Korean Journal of Veterinary Service
• /
• v.45 no.1
• /
• pp.31-38
• /
• 2022

Swine influenza (SI) is an important respiratory disease in pigs and epidemic worldwide, which is caused by influenza A virus (IAV) belonging to the family of Orthomyxoviridae. As seen again in the 2009 swine-origin influenza A H1N1 pandemic, pigs are known to be susceptible to swine, avian, and human IAVs, and can serve as a 'mixing vessel' for the generation of novel IAV variants. To this end, the emergence of swine influenza viruses must be kept under close surveillance. Herein, we report the isolation and phylogenetic study of a swine IAV, A/swine/Korea/21810/2021 (sw21810, H3N2 subtype). BLASTN sequence analysis of 8 gene segments of the isolated virus revealed a high degree of nucleotide similarity (94.76 to 100%) to porcine strains circulating in Korea and the United States. Out of 8 genome segments, the HA gene was closely related to that of isolates from cluster I. Additionally, the NA gene of the isolate belonged to a Korean Swine H1N1 origin, and the PB2, PB1, NP and NS genes of the isolate were grouped into that of the Triple reassortant swine H3N2 origin virus. The PA and M genes of the isolate belonged to 2009 Pandemic H1N1 lineage. Human infection with mutants was most common through contact with infected pigs. Our results suggest the need for periodic close monitoring of this novel swine H3N2 influenza virus from a public health perspective.

• Korean Journal of Food Science and Technology
• /
• v.50 no.5
• /
• pp.480-485
• /
• 2018

In this study, to identify single nucleotide polymorphisms (SNPs) associated with meat quality in Berkshire pigs, we performed RNA sequencing. A non-synonymous SNP (nsSNP) in the Complement component 9 (C9) gene was identified, and the association between meat quality traits and the C9 genotype was analyzed. The nsSNP in the C9 gene was located at c.942 G>T. In the dominant model, significant associations were observed between the SNP and meat quality traits such as CIE L, collagen content, moisture level, and $pH_{24h}$, whereas in the co-dominant model, significant associations were observed between the SNP and CIE L, collagen content, and protein content. In the recessive model, a significant association between the C9 genotype and the collagen content was observed. In addition, we identified the significant relationship between the C9 genotype and meat quality according to sex. These results indicate that the C9 SNP can be used as a genetic marker for improving pork quality.

• Asian Pacific Journal of Cancer Prevention
• /
• v.13 no.9
• /
• pp.4363-4368
• /
• 2012

The epidermal differentiation complex (EDC) contains a large number of gene products which are crucial for the maturation of the human epidermis and can contribute to skin diseases, even carcinogenesis. It is generally accepted that activation of oncogenes and/or inactivation of tumor suppressor genes play pivotal roles in the process of carcinogenesis. Here, NICE-3, a novel EDC gene, was found to be up-regulated in human hepatocellular carcinoma (HCC) by quantitative real-time RT-PCR. Furthermore, overexpression of exogenous NICE-3 by recombinant plasmids could significantly promote cell proliferation, colony formation and soft agar colony formation in Focus and WRL-68 HCC cell lines. Reversely, NICE-3 silencing by RNA interference could markedly inhibit these malignant phenotypes in YY-8103 and MHCC-97H cells. Moreover, cell cycle analysis of MHCC-97H transfected with siRNA by flow cytometry showed that NICE-3 knockdown may inhibit cell growth via arrest in G0/G1 phase and hindering entry of cells into S phase. All data of our findings indicate that NICE-3 may contribute to human hepatocellular carcinoma by promoting cell proliferation.

• Journal of the Korean Society of International Agriculture
• /
• v.30 no.4
• /
• pp.277-284
• /
• 2018

Meat quality has always been one of the most important factors that controls the choice of pork consumers and is of great interest in the pig industry. In this study, we identified a single nucleotide polymorphism (SNP) in the enoyl-CoA delta isomerase 2 (ECI2) gene in Berkshire pigs (n = 430) by analyzed the association between the SNP and meat quality traits. The non-synonymous SNP in the ECI2 gene is located at c.608 C > G and resulted in an amino acid change from threonine to serine. Significant associations between the SNP and meat quality traits, such as redness (CIE a), and the $pH_{24hr}$, were revealed in both the dominant and co-dominant models, whereas carcass weight, drip loss, and fat content and moisture content were significantly associated only with the dominant model. In barrow, the SNP was significantly associated with CIE a, drip loss, and $pH_{24hr}$, whereas in gilt, only a significant relationship with moisture content was observed. GG genotype pigs had a higher $pH_{24hr}$ and lower drip loss. Because $pH_{24hr}$ and drip loss are considered the most important meat quality traits, GG genotype pork is high-quality meat compared to that from other genotypes. In conclusion, the SNP in the ECI2 gene is significantly associated with several meat quality traits. These traits and could be as genetic markers in molecular breeding programs for improving meat quality.

• Proceedings of the Korean Society for Applied Microbiology Conference
• /
• 1986.12a
• /
• pp.514.3-515
• /
• 1986

λP$_{L}$ promoter와 Infiuenza virus의 NS1 Structural gene이 있는 pASl EH801 plasmid를 E. coli host N5' 과 AR120에 각각 transformation하여 온도와 nalidixic acid로 각각 induction 하여 보았다 N5151의 경우, O.D.600 1.2에서 42$^{\circ}C$로 induction하였을 때 maximum productivity를 보였으며 AR120의 경우는 O. D. 600 1.2, 37$^{\circ}C$, 40$\mu\textrm{g}$ nalidixic acid/$m\ell$ induction 하였을 때 maximum yield를 보여주었다. 이때 pH, DO, temperature, $O_2$%, $CO_2$%를 A/D converter통해 computer에 연결시켜 data acquision을 한 결과, 접종 후 ON-line induction이 가능함을 알 수 있었다.

• Journal of Microbiology and Biotechnology
• /
• v.13 no.4
• /
• pp.598-606
• /
• 2003

Salmonella encodes an enterotoxin (Stn) which possesses biological activity similar to the cholera toxin. Stn contributes significantly to the overall virulence of S. typhimurium in a murine model. The production of Stn is enhanced in a high-osmolarity medium and by contact with epithelial cells. In the present study, the in vitro and in vivo transcriptional regulations of the sin promoter revealed two promoters, P1 and P2. The P1 promoter identified by a primer extension analysis of stn mRNA exhibited a switching mechanism in vivo. Depending on the growth stage, transcription was initiated from different start sites termed $P1_S\;and\;P1_E$. $P1_S$, recognized by RNA polymerase containing ${\sigma}^S(E{\sigma}^S),\;and\;P1_E$, recognized by $E{\sigma}^70$, were activated during the stationary and exponential phases, respectively, while $P1_S\;and\;P1_E$ were both negatively regulated by CRPㆍcAMP and H-NS. Results revealed that $P1_S$ was the responsible promoter activated under a high osmolarity and low pH. The P2 promoter was identified 45 nucleotides downstream from $P1_E$ and negatively controlled by CRPㆍcAMP in vitro. No P2 activity was detected in vivo. The regulation of stn expression monitored using a Pstn::egfp fusion indicated that $E{\sigma}^S$ was required for the induction of stn and various factors were involved in stn regulation inside animal cells.

• Journal of Microbiology and Biotechnology
• /
• v.28 no.6
• /
• pp.997-1006
• /
• 2018

As shown during the 2009 pandemic H1N1 (A(H1N1)pdm09) outbreak, egg-based influenza vaccine production technology is insufficient to meet global demands during an influenza pandemic. Therefore, there is a need to adapt cell culture-derived vaccine technology using suspended cell lines for more rapid and larger-scale vaccine production. In this study, we attempted to generate a high-growth influenza vaccine strain in MDCK cells using an A/Puerto/8/1934 (H1N1) vaccine seed strain. Following 48 serial passages with four rounds of virus plaque purification in MDCK cells, we were able to select several MDCK-adapted plaques that could grow over $10^8PFU/ml$. Genetic characterization revealed that these viruses mainly had amino acid substitutions in internal genes and exhibited higher polymerase activities. By using a series of Rg viruses, we demonstrated the essential residues of each gene and identified a set of high-growth strains in MDCK cells ($PB1_{D153N}$, $M1_{A137T}$, and $NS1_{N176S}$). In addition, we confirmed that in the context of the high-growth A/PR/8/34 backbone, A/California/7/2009 (H1N1), A/Perth/16/2009 (H3N2), and A/environment/Korea/deltaW150/2006 (H5N1) also showed significantly enhanced growth properties (more than $10^7PFU/ml$) in both attached- and suspended-MDCK cells compared with each representative virus and the original PR8 vaccine strain. Taken together, this study demonstrates the feasibility of a cell culture-derived approach to produce seed viruses for influenza vaccines that are cheap and can be grown promptly and vigorously as a substitute for egg-based vaccines. Thus, our results suggest that MDCK cell-based vaccine production is a feasible option for producing large-scale vaccines in case of pandemic outbreaks.

• Journal of Microbiology and Biotechnology
• /
• v.18 no.6
• /
• pp.1024-1032
• /
• 2008

Efficient expression of the Salmonella Typhimurium tdc ABCDEG operon involved in the degradation of L-serine and L-threonine requires TdcA, the transcriptional activator of the tdc operon. We found that the tdcA gene was transiently activated when the bacterial growth condition was changed from aerobic to anaerobic, but this was not observed if Salmonella was grown anaerobically from the beginning of the culture. Expression kinetics of six tdc genes after anaerobic shock demonstrated by a real-time PCR assay showed that the tdc CDEG genes were not induced in the tdcA mutant but tdcB maintained its inducibility by anaerobic shock even in the absence of tdcA, suggesting that an additional unknown transcriptional regulation may be working for the tdcB expression. We also investigated the effects of nucleoid-associated proteins by primer extension analysis and found that H-NS repressed tdcA under anaerobic shock conditions, and fis mutation delayed the peak expression time of the tdc operon. DNA microarray analysis of genes regulated by TdcA revealed that the genes involved in N-acetylmannosamine, maltose, and propanediol utilization were significantly induced in a tdcA mutant. These findings suggest that Tdc enzymes may playa pivotal role in energy metabolism under a sudden change of oxygen tension.

• Horticultural Science & Technology
• /
• v.32 no.6
• /
• pp.879-886
• /
• 2014

This study was conducted to test in vitro the antagonistic effect of composted liquid manure (CLM) against soil-borne turfgrass pathogenic fungi, Rhizoctonia solani AG-2-2 (IIIB) (brown patch), R. solani AG-2-2 (IV) (large patch), and Sclerotinia homoeocarpa (dollar spot) for environmentally friendly turfgrass management. CLMs were collected from 9 livestock excretion treatment facilities around the country including Gunwi (GW), Hapcheon (HC), Hoengseong (HS), Icheon (IC), Iksan (IS), Muan (MA), Nonsan (NS), and Yeoju (YJ). CLMs of IC, GW, and IS showed s ignificant (p < 0.05) mycelium growth inhibition that was 17.8%, 20.4%, and 48.0% against R. solani AG-2-2 (IIIB), R. solani AG-2-2 (IV), and S. homoeocarpa, respectively. A t otal of 110 bacterial isolates were obtained from the CLMs that showed antagonistic effects. Among them, 5, 4, and 10 microbe isolates showed promising antifungal activity against mycelium growth of R. solani AG-2-2 (IIIB), R. solani AG-2-2 (IV), and S. homoeocarpa, respectively. The bacterial isolates ICIIIB60, GWIV70, and ISSH20 effectively inhibited the mycelial growth of three soil-borne turfgrass pathogens. Selected bacterial isolates were identified as Alcaligenes sp., Bacillus licheniformis Ab2, and B. subtilis C7-3 through 16s rDNA gene sequence analysis. Among 5 fungicides, the most compatible fungicide with ICIIIB60, GWIV70, and ISSH20 was tebuconazol, toclofos-methyl and toclofos-methyl, respectively. These findings suggested that CLMs could be effectively used not only as organic liquid fertilizer sources but also as biological control agents for soil-borne turfgrass diseases such as brown patch, large patch, and dollar spot.