• International Journal of Industrial Entomology and Biomaterials
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• v.6 no.1
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• pp.93-97
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• 2003

We describe here the cloning of a cDNA encoding putative calreticulin (CRT) from the silkworm, Bombyx mori. The CRT cDNA comprised of 1,194 bp encoding 398 amino acid residues. B. mori. CRT has a HDEL sequence at the end of the C-domain. The B. morl, CRT showed 88% protein sequence identity to the G. mellonella CRT, 71 % to A. aegypti CRT, and 63% to H. sapiens CRT, Phylogenetic analysis revealed that the deduced amino acid sequences of the B. mori CRT formed a highly inclusive subgroup with other insect CRTs. Northern blot analysis exhibited an expression of the B. mori CRT gene in the fat body, evidencing the fat body as a major site for CRT synthesis.

• Journal of the Korean Wood Science and Technology
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• v.46 no.4
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• pp.402-414
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• 2018

Species identification of wood provides important information for archaeology, restoration of cultural assets, preventing illegal logging, and more. Wood species are usually identified based on their anatomical features with the use of a microscope. However, this method may not be able to distinguish between anatomically similar species or subspecies. To overcome this problem, wood species need to be identified at the molecular level using DNA sequencing. However, unlike living plant cells, wood is difficult to pulverize using a mortar, and DNA extraction from dried wood is challenging. To solve these problems, we propose a pretreatment method in which wood is pulverized using 60-grit sandpaper and hydrated with water for 2 days. Using this method, we were able to stably amplify the rpoB gene from the extracted DNA of Pinus rigida. In addition, sequence analysis of the rpoB gene revealed six single nucleotide polymorphisms (SNPs), which classified the rpoB sequences in the genus Pinus into five groups. Our data indicate that although these SNPs were not suitable for species identification, they can potentially be used to determine the origin of different wood subspecies or individual samples of wood.

• ALGAE
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• v.22 no.1
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• pp.1-10
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• 2007

To gain insights into the phylogenetic relationships of genus Staurastrum and Staurodesmus, we analyzed nuclearencoded small subunit rDNA of 82 strains, and chloroplast atpB gene sequences of 44 strains belonging to three genera (Staurastrum, Staurodesmus, Cosmarium). Excluding the Staurastrum muticum and S. orbiculare, forty five strains of genus Staurastrum formed a well supported clade. It was shown that with no cell wall sculpture and processes, these two species have a strong phylogenetic relationship with genus Staurodesmus. Therefore, it is strongly recommended to transfer Staurastrum without processes and cell wall sculpture into Staurodesmus. S. obsoletus is a taxa that is transferred from Cosmarium. But, from this study, it has shown a phylogenetic relationship with Cosmarium. Therefore, this species is strongly recommended to transfer back to Cosmarium instead of Staurodesmus. As it was studied before, genus Staurastrum has shown monophyletic. Since the genus taurodesmus groups with Cosmarium, they were shown to be polyphyletic.

• Tuberculosis and Respiratory Diseases
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• v.39 no.3
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• pp.242-247
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• 1992

Background: The ability to precisely measure specific mRNA levels by hybridization to complementary DNA probes is an important tool for analyzing the regulation of gene expression. Surfactant proteins have important roles in regulating surfactant metabolism as well as in determing its physical properties. Method: The complete coding regions for rat surfactant protein complementary DNA of surfactant protein B were subcloned into pGem 3Z or 4Z such that either antisense or sense transcripts were obtained by using SP 6 RNA polymerase. Surfactant protein B mRNA was measured by filter hybridization. Results: Equation of standard curve between counts per minute (Y) and surfactant protein B mRNA transcript input (X) was Y=2034.9 X+159.1. Correlation coefficient was 1.0. Couclusions: Filter hybridization assay is suited to situation when rapid, accurate quantitation of multiple samples is required.

• Biomolecules & Therapeutics
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• v.9 no.2
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• pp.131-139
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• 2001

Lamivudine, an oral nucleoside analogue, effectively inhibits hepatitis B virus replication and reduces hepatic necroinflammation in patients with chronic hepatitis B. Although lamivudine has shown a promise in patients with chronic hepatitis B, a long-term data on Korean patients with hepatitis B are lacking. The purpose of this study is to evaluate the effects and safety of 52-week lamivudine therapy in Korean patients with chronic hepatitis B, A total of twenty-nine patients (27 male and 2 female) who had received 100 mg of oral lamivudine daily for 52 weeks were evaluated, retrospectively. The mean age of 29 patients in the study group was 37.7 $\pm$ 8.9 years (range 19-54). Pretreatment HBV PCR and HBsAg were positive in all 29 patients, and HBeAg were positive in 25 patients (86%). The serum HBV DNA of 28 patients (97%) significantly fell to undetectable levels (<5 pg/ml) within 12 weeks of therapy and it remained undetectable in 24 patients (83%) by the end of 52-week therapy (p<0.001). Mean serum ALT levels of 29 patients declined to the normal range within 12 weeks and remained within the normal range during the evaluative period (p<0.05). The proportions of patients with HBeAg seroconversion (loss of HBeAg, development of antibody to HBeAg, and undetectable HBV DNA) were 42% after 52-week therapy. The differences of response to lamivudine therapy in HBeAg- positive and HBeAg-negative patients were negligible (p>0.05). Furthermore, the study showed that pretreatment serum HBV DNA and ALT levels have no effect to the efficacy of lamivudine therapy (p>0.05). Further comparison of lamivudine's efficacy showed that lamivudine is just as efficacious in patients with cirrhosis as without cirrhosis (p>0.05). In conclusion, lamivudine is an effective and safe therapy for the treatment of chronic hepatitis B in Korean patients.

• Journal of Yeungnam Medical Science
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• v.15 no.2
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• pp.237-245
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• 1998

만성 B형 간염 환자에서 Interferon (IFN) 치료 후 혈청 HBeAg 소실 및 anti-HBe의 양전율을 높이고 효율적인 치료의 근거를 알기 위하여 치료 전 간기능검사상 갑자기 상승한 혈청 ALT치를 나타낸 환자군과 그렇지 않은 대조군을 대상으로 하여 IFN을 투여한 군과 IFN 치료없이 정상 HBeAg의 자연 소실을 보인 환자군을 임상적으로 장기간 관찰하고 조사하였다. ALT치가 정상 상한치의 4배 이상 높이 증가되어 3개월 이상 왕복을 보인 40명의 환자(A군)와 ALT치가 정상 상한치의 3배 이하로 증가된 10명(B군)에게 ${\alpha}$-IFN 2b를 매일 300만 단위 피하주사로 3~12개월 주사하였다. 대조군으로는 ALT치가 A군처럼 장승한 45명 (C군)이었으며, IFN 치료없이 평균 2.9년을 관찰하였다. HBeAg/anti-HBe 혈청 양전율은 A군 68%, B군 20%, C군 13%이었으며 IFN 치료 중단 후 1년까지의 HBeAg 재양성율은 A군에서 29%였고 HBeAg이 소실된 A와 B군의 38명중에서 6명에서 HBV DNA가 양성이었다. 6명중 4명은 HBeAg/anti-HBe 양전을 보였으나 HBV DNA 양성이었고 나머지 2명은 HBeAg, anti-HBe 및 HBV DNA (hybridization) 모두 음성이었으나 중합효소연쇄반응검사상 HBV DNA 양성이었다. 이상의 결과를 보면 비록 IFN 치료 후에 HBeAg이 소실되었다가 다시 양성화되더라도 IFN은 단기간내에 혈중 HBeAg이나 DNA가 자연적으로 감소가 될 환자나 그렇지 않은 환자에게도 HBV의 비증식화를 유발하여 도움이 될 것으로 사료된다. 그러나 IFN 투여 후에도 혈중 HBeAg과 DNA 소실에 전혀 도움이 되지 않을 환자 및 HBV 증식 억제효과가 기대되는 HBV 간질환 환자의 조건, IFN 투여량, 기간 등에 대한 계획적이고 체계적인 연구로 더 나은 치료효과를 기대할 수 있으리라 생각된다.

• Journal of Microbiology and Biotechnology
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• v.11 no.5
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• pp.819-824
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• 2001

The principal DNA modification systems of the amino-acid-producing bacteria Corynebacterium glutamicum AS019, Brevibacterium flavum BF4, and B. lactofermentum BL1 was investigated using two approaches; digestion of plasmid DNA isolated from these species TseI and Fnu4HI, and sequence analysis of the putative methyltransferase target sites following the derivatization of DNA using metabisulfite treatment. The C. glutamicum and B. flavum strains showed similar digestion patterns to the two enzymes, indicating that the target for cytidine methyltransferase recognizes 5'-GCSGC-3'(where S is either G or C). Mapping the methylated cytidine sites by bisulfite derivatization, followed by PCR amplification and sequencing, was only possible when the protocol included an additional step eliminating any underivatized DNA after PCR amplification, thereby indicating that the derivatization was not $100\%$ efficient. This may have been due to the high G0C content of this genus. It was confirmed that C. glutamicum AS019 and B. flavum BF4 methylated the cytidine in the $Gm^5CCGC$ sequences, yet there were no similar patterns of methylation in B. lactofermentum, which was consistent with the distinctive degradation pattern seen for the above enzymes. These findings demonstrate the successful application of a modified bisulfite derivatization method with the Corynebacterium species for determining methylation patterns, and showed that different species in the geneus contain distinctive restriction and modification systems.

• Korean Journal of Medicinal Crop Science
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• v.22 no.6
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• pp.489-496
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• 2014

Polygonatum is a genus placed in the family Liliaceae, distributed throughout the Northern Hemisphere and 16 of the species are grown naturally in Korea. In oriental medicine, the rhizomes of Polygonatum have been used as two different medicines, Okjuk (Polygonati odorati Rhizoma) and Hwangjeong (Polygonati Rhizoma). However, it is difficult to identify the morphological and chemical differences between the medicinal groups and thus easy to confuse the one with the other. Therefore, a clear classification standard needs to be established so as to be able to discriminate between them. In the study, the morphological characteristics of the plants, Polygonatum spp., were examined. Then, the differences in SNPs among the DNA sequences of 7 of the Polygonatum spp. and 1 of the Disporum spp. were analyzed by DNA barcoding with rpoC1, rpoB2, matK, and psbA-trnH of the cpDNA region. In the results, three regions, rpoC1, rpoB2, and matK were useful for discriminating the species, P. stenophyllum and P. sibiricum. Furthermore, it was possible to discriminate the individual germplasm within the species by using the combination of the results obtained from rpoB2, rpoC1, and matK.

• Korean Journal of Veterinary Research
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• v.39 no.6
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• pp.1151-1160
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• 1999

This study was conducted to detect the toxoplasma-specific DNA in peripheral blood collected from cats experimentally infected with Toxoplasma gondii (RH strain) and from domiciled cats by B1 gene-base polymerise chain reaction(PCR). The sensitivity of oligonucleotide primer, T-1 & T-2, designed from toxoplasma B1 gene amplification method was compared with parasite detection by mouse inoculation(MI). And also, latex agglutination test(LAT) and indirect fluorescent antibody test(IFAT) were conducted to detect the fluctuation of serum antibodies compared with the detection of toxoplasma by PCR and MI. Toxoplasma B1 gene PCR was shown consistently high sensitivity and the results obtained by PCR agreed completely with those from MI. All blood samples collected before infection with T gondii gave negative results by PCR and MI. Also, toxoplasma Bl gene PCR was not cross reaction with Neospora caninum DNA and normal cat leucocyte as controls. The toxoplasma-specific DNA was detected by PCR in blood of 5 cats experimentally infected with T gondii 6 days after infection and the detection of this specific-DNA was long lasted in blood for 64 days after infection. The detection of toxoplasma-specific DNA by PCR could be identified as few as 10 tachyzoites and the isolation of T gondii by MI could be isolated as few as 1 tachyzoite from tenfold serial dilution of T gondii with normal cat blood, respectively. In healthy domiciled cats, the toxoplasma-specific DNA and the parasite were detected and isolated in blood from 3 of 56(5.3%) cats by both PCR and MI, respectively. In the results of antibody test from the total 56 heads of healthy domiciled cats, the positive rates are 15(26.7%) by LAT and 19(33.9%) by IFAT. These results suggest that PCR detection of toxoplasma can be applied as a sensitive and specific diagnostic and research tool.

• BMB Reports
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• v.44 no.9
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• pp.607-612
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• 2011

Several investigators have shown that CpG-DNA has outstanding effects as a Th1-responsive adjuvant and that its potent adjuvant effects are enhanced by encapsulation with a liposome of proper composition. In this study, we showed that encapsulation with phosphatidyl-${\beta}$-oleoyl-${\gamma}$-palmitoyl ethanolamine (DOPE): cholesterol hemisuccinate (CHEMS) complex enhances the immunostimulatory activity of CpG DNA and the binding of CpG-DNA to TLR9. We also examined involvement of myeloid differentiation protein (MyD88) and NF-${\kappa}B$ activation in liposome-encapsulated CpG-DNA-induced IL-8 promoter activation. In this manuscript, the natural phosphodiester bond CpG-DNA encapsulated by DOPE : CHEMS complex is designated as Lipoplex(O). Importantly, we successfully screened B cell epitopes of envelope protein (E protein) of hepatitis C virus (HCV-E) and attachment glycoprotein G of human respiratory syncytial virus (HRSV-G) by immunization with complexes of several peptides and Lipoplex(O) without carriers. Therefore, Lipoplex(O) is potentially applicable as a universal adjuvant for peptide-based epitope screening and antibody production.