• Journal of Plant Biotechnology
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• v.33 no.4
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• pp.309-313
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• 2006

Discovery of single nucleotide polymorphisms (SNPs), including small insertions and deletions, is one of the hot topics in genetic research. The most common type of sequence variant consists of single base differences or small insertions and deletions at specific nucleotide positions. Significance of SNPs in rice is increasing for genetic research, positional cloning and molecular breeding. $F_2$ 170 lines and $F_3$ 194 lines derived from Sangjuchalbyeo/HR13721-53-3-1-3-3-2-2 Were used for Searching SNP markers related to bacterial blight resistance. Sangjuchalbyeo is susceptible to bacterial blight, but HR13721-53-3-1-3-3-2-2 has Xa1 gene resistant to bacterial blight. Individual lines were inoculated with $K_1$ race of bacterial blight and resistant or susceptible was evaluated after 3 weeks from inoculation. The genotypes of population were analysed by PCR-RFLP for SNP marker developing. The segregation of $F_2\;and\;F_3$ population showed almost 3:1, 1:1 ratio, respectively. Analysis of genotype using SNP marker is capable of confirming resistance for $K_1$ race and genotype through amplifying the gene using 16PFXal primer and digested the PCR product with Eco RV. There were close relation between resistance test for $K_1$ race and SNP marker genotype. Especially, DNA analysis using SNP marker is capable of judging homozygote/heterozygote in $F_2$ population compared with resistant test for Kl race. So, it seems to improve the selection efficiency in disease resistant breeding.

• Korean Journal of Veterinary Research
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• v.40 no.4
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• pp.653-660
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• 2000

In order to study the regional distribution and relative frequency of the immunoreactive endocrine cells in the gastrointestinal tract of the Mongolian gerbil, Meriones unguiculatus, the gastrointestinal tract was divided into 9 portions (cardia, fundus, pylorus, duodenum, jejunum, ileum, cecum, colon and rectum) and immunostained by immunohistochemical (PAP) method using 8 types of specific antisera against cholecystokinin (CCK)-8, gastrin, secretin, pancreatic polypeptide(PP), somatostatin, serotonin, glucagon and insulin. CCK-8-, gastrin-, somatostatin- and serotonin-immunoreactive cells were demonstrated in this study. These immunoreactive cells were found in the gastric gland regions of the pylorus or between parietal and chief cells of the fundus with round to spherical shaped, and in the interepithelial regions of the intestinal tract with spherical to spindle shaped except for jejunum where some of immunoreactive cells were also observed in the intestinal glands with round to spherical shaped. CCK-8-immunoreactive cells were restricted to the pylorus and duodenum with numerous and a few frequency, respectively. Gastrin-immunoreactive cells were restricted to the pylorus with numerous frequency. Similar to those of gastrin-immunoreactive cells, somatostatin-immunoreactive cells were restricted to pylorus with moderate frequency. Serotonin-immunoreactive cells were detected throughout whole gastrointestinal tract except for cardia and cecum with moderate or numerous frequency. However, no secretin-, PP-, glucagon- and insulin-immunoreactive cells were observed in this study. From these results, the appearance type, regional distribution and relative frequency of immunoreactive endocrine cells in the gastrointestinal tract of the Mongolian gerbils were somewhat lowered or restricted compared to those of other mammals and these differences were might be caused by feeding habits and species specification.

• Pediatric Infection and Vaccine
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• v.12 no.2
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• pp.166-177
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• 2005

Purpose : The purpose of this study was to examine the molecular epidemiology and genetic variability of adenovirus(Ad) serotypes Ad1, Ad2, and Ad5 over 14 years in Korea. Methods : A total of 382 adenoviral strains isolated from the nasopharyngeal aspirates of children with lower respiratory tract infections in Seoul, Korea from November 1990 to February 2003 were serotyped by neutralization assay with type-specific antisera. Viral DNAs were extracted from infected cell lysates by the modified Hirt procedure. Genome type(GT) was determined by DNA restriction analysis with 12 restriction enzymess(BamHI, BclI, BglI, BglII, BstEII, EcoRI, HindIII, HpaI, SalI, SmaI, XbaI, and XhoI). To evaluate the genetic relatedness, pairwise comigrating restriction fragments(PCRF) analysis was performed. Results : Of 382 strains, 33 strains(9%) were Ad1, 45 strains(12%) were Ad2, and 24 strains(6%) were Ad5. Eighteen GTs(Ad1p1-Ad1p7, Ad1a, Ad1b, Ad1b1-Ad1b3, Ad1c, Ad1d, Ad1e, Ad1e1, Ad1e2, Ad1f) among Ad1, 24(Ad2p1-Ad2p11, Ad2a, Ad2a1-Ad2a6, Ad2b, Ad2c, Ad2d, Ad2e, Ad2e1-Ad2e3) among Ad2, and 10(Ad5p1, Ad5p2, Ad5a, Ad5a1-Ad5a7) among Ad5 strains were identified. One or two strains of the vast majority of GTs were isolated during the study period while a few GTs were identified sporadically with more than 2 strains. It is notable that some GTs such as Ad1p5 and Ad5a1 appeared in cluster during a short period. In analysis of genetic relatedness, the degree of PCRFs(pairwise comigrating restriction fragments) for Ad1 varied from 79 to 99%, for Ad2, 82 to 99%, and for Ad5, 85 to 99%. Conclusion : This study established the comprehensive nomenclature systems of Ad1, Ad2, and Ad5. Diverse GTs identified in this study have crucial implications in the genomic diversity and epidemiological characteristics of Ad1, Ad2, and Ad5.

• Korean Journal of Food Science and Technology
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• v.40 no.4
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• pp.424-429
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• 2008

To investigate the acid tolerance characteristics of the acid-resistant mutant, Leuconostoc paramesenteroides P-200, as a kimchi starter, this study examine proton permeability, ATPase activity, glycolysis activity, $Mg^{2+}$ release, and membrane fatty acid composition, and compared the data to that of its wild-type, L. paramesenteroides LP-W. In the proton permeability experiment, the LP-W and P-200 strains' average maximum half-time $(t_{1/2})$ values for pH equilibration through the cell membrane were approximately 5.7 and 9.3 min in 150mM KCl solution, and 4.2 and 8.3 min in 3% NaCl solution, respectively. Their values and pH levels for maximal specific ATPase activity showed that P-200 had greater activity than LPW. And the results of pH-dependent glycolysis activity showed that P-200 had greater activity than LP-W. Furthermore, after 2 hr at pH 4.0, LP-W and P-200 had percent magnesium release values of approximately 12% and 34%, respectively. A comparison of their membrane fatty acid compositions indicated that C18 and cyclo-C19 were the major different fatty acids between the two strains, and their contents of C18 and cyclo-C19 were 2.5% and not detected, respectively, in LP-W, and 6.4% and 11.4%, respectively, in P-200. These results indicate that the P-200 strain has significantly improved acid tolerance as compared to its wild type, LP-W.

• Journal of Life Science
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• v.26 no.3
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• pp.275-281
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• 2016

Glycan modification is important in pharmaceutical industry. Especially, sialic acid affects the bioactivity and stability of medicine. Milk of pig has been used as bioreactor to produce various pharmaceutical proteins. Therefore, it is necessary to modify the glycan chain in pig mammary grand. β-1,4-N-Acetylglucosaminyltransferase A (pMGAT4A) is one of the essential enzymes for increase of sialic acid content, but pig MGAT4A is unclear. In this study, the pMGAT4A was identified and characterized. The pMGAT4A has 1638 nucleotides encoding 535 amino acids and type II membrane topology, which is one of the common features in many glycosyltransferases. The gene was strongly expressed in liver and mammary gland, whereas was weakly expressed in small intestine, stomach and bladder. For functional test, HA-tagged MGAT4A was over-expressed in porcine kidney (PK-15) cell line. Forced expression of pMGAT4A gene was identified by qPCR, and we identified that pMGAT4A is located in Golgi complex by co- staining with HA antibody and BODIPY TR ceramide. In addition, we identified the increase of mannose-β-1,4-N-acetylglucosamine structure by ELISA and immunofluorescence using Datura stramonium agglutinin (DSA), which recognizes mannose-β-1,4-Nacetylglucosamine. Through the specific activity analysis, we showed that pMGAT4A modified bi-antennary to tri-antennary. This event affects sialic acid content. Therefore, we thought that over-expression of pMGAT4A will be necessary in pig mammary grand for improved medicine.

• Microbiology and Biotechnology Letters
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• v.26 no.2
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• pp.102-109
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• 1998

To investigate the increased acid tolerance of a acid-resistant mutant Leuconostoc mesenteroides M-100 improved as a kimchi starter, proton permeability, ATPase acitivity, glycolysis activity, $Mg^{2+}$ releasement, and membrane fatty acid composition were studied and comprised with its wild type Leuconostoc mesenteroides Mw. In the proton permeability experiment, the mininum values of the average half time ($t_{1/2}$) of pH equilibration through the cell membrane of the Mw and the M-100 were about 8.6 min and 9.2 min in 150 mM KCI solution, respectivily. In the 3% NaCl solution, the $t_{1/2}$ values of the Mw and the M-100 were 6min and 8.6 min, respectivily. The values and pHs of maximal specific activities of ATPase originated from the Mw and the M-100 were 0.6U at pH 5.5 and 0.8U at pH 5.5, respectivily. The result of pH dependence of glycolysis showed that the M-100 had higher activities than that of Mw except at pH 5.0. The releases of magnesium from the Mw and the M-100 were observed about 36.5% and 13% at pH 4.0 after 2 hours, respectivily. The results of comparison of membrane fatty acid composition of the Mw with the M-100 showed that $C_{12}$, $C_{14}$, $C_{18:1}$, and $C_{19:0cyclo}$ were major different fatty acids between two strains and the content of $C_{18:1}$, and $C_{19:0cyclo}$ were 23.4%, 10.2% in the Mw and 15.1%, 12.2% in the M-100. These results indicated that acid tolerance of the M-100 was significantly improved in comparison with its wild type Mw.

• Journal of Food Hygiene and Safety
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• v.31 no.2
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• pp.113-118
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• 2016

Proprotein convertase subtilisin/kexin type 9 (PCSK9), is a protein mainly secreted by a liver. The PCSK9 plays an important role in low density lipoprotein (LDL) metabolism acting as a repressor of LDL receptor through transportation of the LDLR to the lysosome for degradation. Thus, the PCSK9 inhibitor suppresses PCSK9-regulated degradation of the LDL receptor as a LDL-lowering medicine. However, little is known about the role of PCSK9 in the reproductive system. Therefore, in the present study, we investigated Pcsk9 expression in male reproductive tracts including penises, prostates and testes using rats in response to their diets between a normal diet and a high-fat diet with cholesterol. Based on our previous study, the high-fat diet elevates concentration of total cholesterol and LDL in serum whereas it reduces the concentration of plasma high density lipoprotein (HDL). In addition, it dramatically affects to morphological changes of the male reproductive organs. Consistent with these results, the expression of Pcsk9 was substantially decreased in the penile tissues (P < 0.001) from rats fed a high fat diet as compared to a normal diet. Moreover, it slightly reduced in the prostate and testes (P < 0.05) of rats in response to a high fat diet. Localization of Pcsk9 was predominantly detected in urethral epithelium of penises, cylinder-shaped cells of prostate glands, and spermatogonia, spermatocytes and spermatid of testes of rats. Collectively, results of current study provide invaluable insights into the Pcsk9 gene with respect to its tissue- and cell-specific expression by a high fat diet with cholesterol.

• Korean Journal of Microbiology
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• v.48 no.2
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• pp.109-115
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• 2012

Helicobacter pylori is an important factor of chronic gastritis, digestive ulcer, and stomach cancer. CagL, a virulence factor of H. pylori, is well-known as a pilus protein which acts as adhesion to host cell and a component of Type 4 secretion system. In this study, we evaluated the protective response of recombinant CagL protein (rCagL) using Mongolian gerbil animal model for H. pylori infection. The cagL gene was cloned from 26695 H. pylori followed by over-expression and purification of the protein in E. coli. Mongolian gerbils were immunized with rCagL protein mixed with aluminum adjuvant via intramuscular injections once a week during 4 weeks. At a week after the last immunization, the Mongolian gerbils were administrated with H. pylori 7.13 strain into the stomach and sacrificed to measure antibody titer on rCagL by ELISA and bacterial colonization in the stomach, and to examine the histopathological changes and cytokine expression at 6 week after challenge. Antibody titers on recombinant protein were significantly increased from a week after the first immunization. There was no significant change of the number of bacterial colony between control group and immunized group. The relative stomach weight was significantly decreased in immunized group, but the significant change of histopathological assessment was not observed in the stomach. Cytokine expression such as IL-$1{\beta}$ and KC also was not significantly different between control and immunized groups. These results indicate that rCagL could effectively induce the formation of the specific IgG antibodies. However, bacterial colonization and histopathological lesions could not be inhibited by the immunization in the stomach, indicating not enough protection against H. pylori infection. We consider that along with CagL other adequate antigens could be needed stimulating immune response and inducing protective effects against gastric disease, and also a better adjuvant could be considered.

• Journal of the korean academy of Pediatric Dentistry
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• v.30 no.2
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• pp.217-228
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• 2003

Co-ordinate growth of the brain and skull is achieved through a series of tissue interactions between the developing brain, the growing bones of the skull and the sutures that unite the bones. Craniosynostosis, the premature fusion of cranial sutures, presumably involves disturbance of these interactions. Bmp2, one of bone morphogenetic proteins (Bmps), is involved in the regulation of the shapes of individual bones and the relative proportions of the skeleton. Mutations in the homeobox gene Msx2, known as a downstream gene of Bmp, cause Boston-type human craniosynostosis. The phenotype of Dlx5 homozygote mutant mouse presents craniofacial abnormalities including a delayed ossification of calvarial bone. These facts suggest important roles of Bmp2, Msx2 and Dlx5 genes in the cranial bone growth and suture morphogenesis. To elucidate the function of these molecules in the early morphogenesis of mouse cranial sutures, we first analyzed by in situ hybridization the expression of Bmp2(E15-18), Msx2 and Dlx5 genes in the developing sagittal suture of calvaria during the embryonic stage. Bmp2 mRNA was intensely expressed in the osteogenic fronts and also at the low level in the periosteum of parietal bones during embryonic stage, Msx2 mRNA was intensely expressed in the sutural mesenchyme and mildly expressed in the dura mater during the embryonic stage. Dlx5 mRNA was intensely expressed osteogenic fronts and parietal bones. To further examine the role of Bmp signaling in cranial suture, we did in vitro experiments in E15.5 mouse calvarial explants. Interestingly, implantation of Bmp2-soaked beads onto the osteogenic fronts after 48 hours organ culture resulted in the increase of the tissue thickness and cell number around Bmp2 beads, compared to BSA control beads. In addition Bmp2 induced etopic expressions of Msx2 and Dlx5 genes. On the other hand, overexpression of FGF2 did not induce the expression of Msx2 and Dlx5. Taken together, these data indicate that Bmp2 signaling molecule has a important role in regulating the cranial bone growth and early morphogenesis of cranial suture. We also suggest that Bmp signaling is involved in all the stages of osteogenesis of cranial bones and the maintenance of cranial suture by regulating Msx2 and Dlx5 genes, and that Msx2 and Dlx5 genes are specific transcription factors of Bmp signaling pathway.

• Development and Reproduction
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• v.11 no.3
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• pp.187-193
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• 2007

Ethane 1,2-dimethane sulfonate(EDS), a Leydig cell specific toxicant, has been widely used to create the reversible testosterone withdrawal rat model. Though the maintenance of epididymal structure and function is highly dependent on the testosterone secreted from testis, its derivatives, dihydroxytestosterone(DHT) and estrogen, might have crucial roles. The aim of present study was to monitor the expression patterns of sex steroid receptors, cytochrome P450 aromatase(P450arom) and $5{\alpha}$-reductase in the rat epididymis up to 7 weeks after EDS injection. Adult male rats($350{\sim}400g$) were injected with a single does of EDS(75 mg/kg i.p.) and sacrificed on weeks 0, 1, 2, 3, 4, 5, 6 and 7. The transcriptional activities of the target genes were evaluated by semi-quantitative RT-PCRs. The transcript level of estrogen receptor alpha($ER{\alpha}$) in EDS group was significantly higher than control level on week 1(P<0.01). After week 2, there was no significant difference in $ER{\alpha}$ levels between EDS group and control. The transcript level of estrogen receptor beta($ER{\beta}$) in EDS group was significantly higher than control level on week 1(P<0.05), lowered on weeks 2 and 3(P<0.05 and P<0.01, respectively), fluctuated during weeks 4 and 6, and elevated on week 7(P<0.05). The androgen receptor (AR) message levels increased significantly week 2(P<0.01), then returned to control level on week 3. In contrast, expression of cytochrome P450 aromatase(P450arom) decreased sharply during weeks $1{\sim}3$(P<0.01 on weeks 1 and 2; P<0.05 on week 3), then went back to control level on week 4. The mRNA level of $5{\alpha}$-reductase type 2($5{\alpha}$-RT2) increased significantly on week 4(P<0.01), then returned to control level. The present study indicated that EDS administration could induce reversible alterations in the transcriptional activities of sex steroid hormone receptors and androgenconverting enzymes in rat epididymis. EDS injection model will be useful to clarify the regulation mechanism of mammalian epididymal physiology.