• 한국패류학회지
• /
• 제15권1호
• /
• pp.13-20
• /
• 1999

A horizontal starch gel electrophoresis for enzyme proteins extracted from three Korean species and one Chinese species of Semisulcospira was carried out in order to elucidate their genetic relationships. A total of 10 enzymes were employed in three different of buffer systems. Two loci from each enzyme of GAPDH, GOT, ICDH, IDH and PEP(VL); three loci from each of three enzymes, EST, PEP(LGG) and PGDH; and five loci from GPI were observed. Most of the loci in three pleurocerid species employed showed homozygous monomorphic banding patterns and some of them were specific as genetic markers between two different species. However, EST-2, PEP(LGG-3) and PGDH-1 loci in Korean S. libertina and PEP(LGG-3), PGM-1 and PGM-2 loci in Chinese S. libertina showed polymorphic banding patterns. Three Korean Semisulcospira species including S. libertina were more closely clustered in a dendrogram within the range of genetic identity values of 0.818-0.936, and these clusters were lineated with Chinese S. libertina at the value of 0.621.

• Journal of Microbiology and Biotechnology
• /
• 제20권12호
• /
• pp.1769-1771
• /
• 2010

The envelope glycoprotein E2 of hepatitis C virus (HCV) binds to various cell surface receptors for viral infection. We performed biopanning against this protein and selected peptides from phage display peptide libraries. Two short peptides, pep7-1 and pep12-1, were selected and their ability to inhibit the infection process was investigated. When pep7-1 was present, the infectivity of HCV particles in cell culture was notably decreased. This decrease was demonstrated by Western blot analysis, immunofluorescence assay, and reverse transcription PCR assay. However, pep12-1 showed little inhibitory effect on HCV infection.

• 한국식품과학회지
• /
• 제32권6호
• /
• pp.1331-1335
• /
• 2000

김치 젖산균인 WL6는 API kit 또는 Biolog system방법에 의하여 동정한 결과, Leuconostoc mesenteroides ssp. cremoris, Leu. mesenteroides ssp. dextranicum 또는 Lactobacillus bifermentans로 나타나 동정되지 않았다. 그러나, pepN gene과 16S rRNA gene으로부터 2개의 specific-sequence primer set을 제조하여 PCR 방법으로 증폭한 후에 표준균주들과 비교한 결과, WL6는 Lactobacillus bifermentans로 추정되었다.

• 한국식물병리학회:학술대회논문집
• /
• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
• /
• pp.69.1-69
• /
• 2003

RSH (relA/spoT homolog) has been known to determine the level of guanosine tetraphosphate (ppGpp) and guanosine pentaphosphate (pppGpp), which are the effector nucleotide of the prokaryotic stringent response and also play a role in antibiotic production and differentiation in Streptomyces species but not a little in eukaryotic organism, especially in plant. Salicylic acid (SA), a critical signal molecule of establishing systemic acquired resistance (SAR), could induce SAR in Pepper (Capcicum annuum) against Phytophthora capsici. And the extent of SAR induction was in proportion to the dosage of SA (or BTH). Suppression subtractive hybridization (SSH), a PCR-based method for cDNA subtraction, was carried out between SA-treated and non-SA-treated pepper leaves to isolate genes which may be responsible for defense signaling against pathogens. Early upregulated gene was selected from reverse northern and kinetics of SSH-genes transcripts in SA-treated pepper leaves upon SA treatment. Full-length cDNA of the gene (PepRSH； Pepper RelA / SpoT homolog) had an open reading frame (ORF) of 2166 bp encoding a protein of 722 amino acids and a significant homology with (p)ppGpp phosphohydrolase or synthetase. Genomic DNA gel blot analysis showed that pepper genome has at least single copy of PepRSH. PepRSH transcripts was very low in untreated pepper leaves but strongly induced by SA and methyljasmonic acid (MeJA), indicating that PepRSH may share common SA and MeJA-mediated signal transduction pathway Functional analysis in E. coli showed PepRSH confers phenotypes associated with (p)ppGpp synthesis through a complementation using active site mutagenesis.

• Journal of Microbiology and Biotechnology
• /
• 제16권3호
• /
• pp.457-464
• /
• 2006

Three extracellular proteases (Vpr, peptidase T, and subtilisin) were identified from the culture supernatant of Bacillus subtilis KCTC 3014. All the proteins were partially purified as a mature form by using a DEAE-cellulose ion-exchange column chromatography. Their activities were determined by using zymography and densitometry. The relative molecular masses of Vpr and peptidase T (PepT) were determined to be 68 and 48 kDa by SDS-PAGE and zymography, respectively. However, subtilisin formed a 'binding mode' at the top of the separating gel. After denaturation by boiling at $100^{\circ}C$ for 5 min, its molecular mass was determined to be 29 kDa, whereas its activity was lost. The optimal pH of Vpr, PepT, and subtilisin were 9.0, 6.0-7.0, and 7.0-8.0, respectively. The optimal temperature of Vpr, PepT, and subtilisin was 40, 50, and $40^{\circ}C$, respectively. Inhibitor test revealed that Vpr and subtilisin were serine proteases and that PepT was a metalloprotease. Interestingly, we found that Vpr showed no enzyme activity on a 2DE zymogram gel. Three genes, vpr, pepT, and apr (encoding subtilisin protein), were cloned and their nucleotide and deduced amino acid sequences were determined.

• 한국미생물·생명공학회지
• /
• 제34권1호
• /
• pp.35-39
• /
• 2006

높은 C4 대사활성을 보이는 반추위미생물이 가지는 포도당 발효대사 조절양식의 한가지를 대장균에서 모사하였다. 대장균은 glycolytic condition에서는 phosphoenolpyruvate(PEP) ${\leftrightarrow}$ oxaloacetate(OAA)간 반응을 phosphenolpyruvate carboxylase(PPC)에 의해, gluconeogenetic condition에서는 phosphoenolpyruvate carboxykinase(PCK)에 의해 촉매하도록 조절한다. 반면 반추위미생물은 glycolytic condition에서 PCK를 통하여 반응이 촉매된다. 이러한 조절양식의 차이점이 C4 대사활성에 미치는 영향을 조사하기 위하며 ppc가 돌연변이되고 대신 인위적으로 PCK를 발현할 수 있는 대장균을 제조하였다. 이렇게 PEP-OAA간 대사조절이 변이된 대장균 K12 ppc-/pck+는 야생형 K12보다 2.5배의 높은 C4대사활성을 보였다. 대장균에서의 C4 대사생리를 증가시키는 연구는 대사공학을 이용한 여러가지 유용물질(i.e. 숙신산, ALA)생산에 응용하기 위한 기초자료로 활용될 수 있을 것으로 기대된다.

• Asian Pacific Journal of Cancer Prevention
• /
• 제16권2호
• /
• pp.571-578
• /
• 2015

Prostate cancer is the most common cancer in men. In this study, we investigated immune responses of cytotoxic T lymphocytes (CTLs) against TRAMP-C2 prostate cancer cells after activation by dendritic cells (DCs) loaded with TRAMP-C2 freeze-thaw antigen and/or PEP-3 peptide in vitro. Bone marrow-derived DC from the bone marrow of the C57BL/6 were induced to mature by using the cytokine of rhGM-CSF and rhIL-4, and loaded with either the freeze-thaw antigen or PEP-3 peptide or both of them. Maturation of DCs was detected by flow cytometry. The killing efficiency of the CTLs on TRAMP-C2 cells were detected by flow cytometry, CCK8, colony formation, transwell migration, and wound-healing assay. The levels of the IFN-${\gamma}$, TNF-${\beta}$ and IL-12 were measured by enzyme-linked immunosorbent assay (ELISA). Compared with the unloaded DCs, the loaded DCs had significantly increased expression of several phenotypes related to DC maturation. CTLs activated by DCs loaded with freeze-thaw antigen and PEP-3 peptide had more evident cytotoxicity against TRAMP-C2 cells in vitro. The secretion levels of IFN-${\gamma}$, TNF-${\beta}$ and IL-12, secreted by DCs loaded with antigen and PEP-3 and interaction with T cells, were higher than in the other groups. Our results suggest that the CTLs activated by DCs loaded with TRAMP-C2 freeze-thaw antigen and PEP-3 peptide exert a remarkable killing efficiency against TRAMP-C2 cells in vitro.

• Clinical and Experimental Reproductive Medicine
• /
• 제24권1호
• /
• pp.51-56
• /
• 1997

Recently, through the development of the primer extension preamplification(PEP) method which amplifies the whole genome, simultaneous multiple DNA analysis has become possible. Whole genome from each single cell can be amplified using 15 base oligonucleotide random primer. The greatest advantage of PEP-PCR is the ability to investigate several loci simultaneously and confirm results by analysing multiple aliquots for each locus. This technique led to the development of preimplantation genetic disease diagnosis using blastomere from early embryo, sperm, polar body and oocyte. In this study, we applied PEP-PCR in 20 cases of single amniocyte and 20 cases of single chorionic villus cell for the clinical application of the prenatal and preimplantational genetic diagnosis. We analysed 7 gene loci simultaneously which are 46, 47 exons related to dystrophin gene, two VNTR (variable number tandem repeat) markers using 5'dysIII, 3'CA related to dystrophin gene and DYZ1, DYZ3, DYS14 regions on chromosome Y. In all the tests, 97.5% of PEP-PCR amplifications with single cells were successful. We obtained 38/40 (95%) accuracy in gender determination through chromosome analysis comparison. Therefore, these results have significant implications for a sperm or oocyte analysis and prenatal or preimplantational genetic diagnosis.

• Journal of Preventive Medicine and Public Health
• /
• 제50권3호
• /
• pp.210-216
• /
• 2017

Objectives: One way to prevent deaths due to rabies is the timely utilization of post-exposure prophylaxis (PEP). Therefore, in addition to an understanding of the epidemiological distribution of animal bites, it is necessary to explore the factors leading to delays in PEP initiation. Methods: This cross-sectional study was conducted in Iran in 2011, and included 7097 cases of animal bites recorded at the Rabies Treatment Center of the Shiraz University of Medical Sciences using the census method. Logistic regression was used to identify factors associated with delays in PEP. Results: Among the patients studied, 5387 (75.9%) were males. The prevalence of animal bites in Fars province was 154.4 per 100 000 people. Dogs were the most frequent source of exposure (67.1%), and the most common bitten part of the body was the hands (45.5%). A delay in the initiation of PEP was found among 6.8% of the studied subjects. This delay was more likely in housewives (odds ratio [OR], 4.66; 95% confidence interval [CI], 2.12 to 10.23) and less likely in people with deep wounds (OR, 0.65; 95% CI, 0.43 to 0.97). Conclusions: Although all animal bite victims received complete PEP, in some cases, there were delays. Further, the type of animal involved, the depth of the bite, and the patient's occupation were the major factors associated with a delay in the initiation of PEP for rabies prevention.

• 한국약용작물학회지
• /
• 제12권1호
• /
• pp.1-9
• /
• 2004

200종의 식물추출물을 대상으로 PEP 저해활성을 측정한 결과, 81종의 식물추출물은 $10{\sim}30%$ 범위의 저해율을, 28종의 식물추출물은 10%이하의 저해율을 나타내어 총 200종의 식물추출물 중 약 55% 정도의 식물추출물이 30%이하의 낮은 PEP 저해활성을 나타내었다. 반면에, Prunus mume (매화나무) 잎 (95.3%), 노루발과 (Pyrolaceae)의 Pyrola japonica (노루발) 잎과 줄기 (93.2%), 물레나물과 (Hypericaseae)의 Hypericum ascyron (물레나물) 지상부 (90.2%), 범의귀과(Saxifragaceae)의 Astilbe chinensis var. typica (노루오줌) 지상부 (90.1%), 보리수나무과 (Elaeagnaceae)의 Elaeagnus umbellata (보리수나무) 잎과 줄기 (90.1%)등 5종의 추출물들이 5 ppm에서 PEP에 대하여 90% 이상의 강한 저해활성을 나타내었다.