• BMB Reports
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• v.39 no.1
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• pp.55-60
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• 2006

We describe a phage display strategy, based on the differential resistance of proteins to denaturant-induced unfolding, that can be used to select protein variants with improved conformational stability. To test the efficiency of this strategy, wild-type and two stable variants of ${\alpha}_1$-antitrypsin (${\alpha}_1AT$) were fused to the gene III protein of M13 phage. These phages were incubated in unfolding solution containing denaturant (urea or guanidinium chloride), and then subjected to an unfavorable refolding procedure (dialysis at $37^{\circ}C$). Once the ${\alpha}_1AT$ moiety of the fusion protein had unfolded in the unfolding solution, in which the denaturant concentration was higher than the unfolding transition midpoint ($C_m$) of the ${\alpha}_1AT$ variant, around 20% of the phage retained binding affinity to anti-${\alpha}_1AT$ antibody due to a low refolding efficiency. Moreover, this affinity reduced to less than 5% when 10 mg/mL skimmed milk (a misfolding-promoting additive) was included during the unfolding/refolding procedure. In contrast, most binding affinity (>95%) remained if the ${\alpha}_1AT$ variant was stable enough to resist unfolding. Because this selection procedure does not affect the infectivity of M13, the method is expected to be generally applicable to the high-throughput screening of stable protein variants, when activity-based screening is not possible.

• Journal of the Microelectronics and Packaging Society
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• v.13 no.2 s.39
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• pp.15-19
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• 2006

The resistance switching characteristics of amorphous $SiO_2$ and poly-crystalline $TiO_2$ were investigated. Both films exhibit well defined switching characteristics with low and high resistance states. From I-V curve analyses, it was found that the low resistance states of both films obey an ohmic conduction mechanism and the high resistance states show generation of a Schottky potential barrier. Regarding the mechanism for resistance switching of the binary oxide, it is suggested that the generation and annihilation of potential barriers accounts for the changes to the high resistance state and low resistance state, respectively. The device operation characteristic parameters such as reset and set voltages of $TiO_2$ are distinctly smaller than those of $SiO_2$, indicating that the values are related to the dielectric constant.

• Parasites, Hosts and Diseases
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• v.37 no.4
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• pp.257-263
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• 1999

Identification of the genes responsible for the recovery of virulence in brain-passaged Acanthamoeba culbertsoni was attempted via mRNA differential display polymerase chain reaction (mRNA DD-PCR) analysis. In order to identify the regulatory changes in transcription of the virulence related genes by the brain passages, mRNA DD-PCR was performed which enabled the display of differentially transcribed mRNAs after the brain passages. Through mRNA DD-PCR analysis. 96 brain-passaged amoeba specific amplicons were observed and were screened to identify the amplicons that failed to amplify in the non-brain-passaged amoeba mRNAs. Out of the 96 brain-passaged amoeba specific amplicons, 12 turned out to be amplified only from the brain-passaged amoeba mRNAs by DNA slot blot hybridization. The clone, A289C, amplified with an arbitrary primer of UBC ＃289 and the oligo dT$_{11}$-C primer, revealed the highest homology (49.8％) to the amino acid sequences of UPD-galactose lipid transferase of Erwinia amylovora, which is known to act as an important virulence factor. The deduced amino acid sequences of an insert DNA in clone A289C were also revealed to be similar to cpsD, which is the essential gene for the expression of type III capsule in group B streptococcus. Upregulated expression of clone A289C was verified by RNA slot blot hybridization. Similar hydrophobicity values were also observed between A289C (at residues 47-66) and the AmsG gene of E. amylovora (at residues 286-305: transmembrane domains). This result suggested that the insert of clone A289C might play the same function as galactosyl transferase controlled by the AmsG gene in E. amylovora.a.

• Parasites, Hosts and Diseases
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• v.35 no.3
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• pp.203-210
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• 1997

The difrrrenlial display reverse transcription polymerase chain reaction (DDRT-PCR) aniilysis roils performed to identify the pathogellir strain specific amplicons. mRNAs were purified from the trophozoites of the pathogenif strain YS-27 and the non-pathogenic strain S 16. respectively. Three kinds of rirsl stranded rDNAs were reverse transcribed from the mRNAs by one base anchored oligo-dT 11M (M: A. C, or G) primers. Each cDNA lemplatr was used for DDRT-PCK analysis. A total of 144 pathogenic strain specific amplicons was observed in DDRT-PCR analysis using primer combinations of the 11 arbitrary primers and the 3 one base anchored oli해-dT11M primers. Of these 31 amplit'tons were verified as the amplirons amplified only from the mRNAs of the pathogenic strain by DNA slots biol llybridizatioil. Furthel cklaracleization of the 31 pathogenic strain sprcifil amplicons by DNA slot blot hybridlnation analysis using biotin labeled Probes or the PCR amplified DNA of rysteine proteinase genes revealed that 21 of them were amplliried from the maNAs of the cysteine proteinase genes. Four randomly selected amplirons out of the rest 10 amplirons were used fur screening of cDNA library followed by immunoscreening and all of them were turned outs to be amplified from the mRNA.

• Journal of the Korean Crystal Growth and Crystal Technology
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• v.33 no.5
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• pp.187-190
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• 2023

In this study, antibacterial glasses were developed by the addition of CaO in ZnO-Na₂O-B₂O₃-SiO₂ glass system. The effect of the addition of CaO on the thermal properties, dissolution properties, surface zeta potential and antibacterial activity were analyzed. Differential thermal analysis (DTA) analysis was performed to analyze the thermophysical properties of 30ZnO-xCaO-20Na₂O-30B₂O₃-(20-x)SiO₂ (x = 0, 2, 4, 6, 8, 10 mol%). It was confirmed that the glass transition temperature decreased as the CaO content increased. The amount of released Zn²⁺ ions and surface zeta potential of glass samples increased with increasing CaO concentration. For these reasons, the antibacterial activity was dramatically improved. By the addition of CaO, we could successfully develop an antibacterial glass with 99.9 % antibacterial activity against both Escherichia coli and Staphylococcus aureus.

• Applied Chemistry for Engineering
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• v.18 no.1
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• pp.77-83
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• 2007

It is essential that second order nonlinear optical materials have low optical propagation losses in the wavelengths of second harmonic generation for practical applications in waveguides. Three dipolar chromophores substituted with nitro, cyano, and alkyl sulfone as an electron withdrawing group were prepared. The UV-Vis absorption spectra of the cyano and alkylsulfone chromophores showed a blue-shift compared to the nitro chromophore. The introduction of oxadiazole segment in the chromophore structure led to similar spectral shift. The blue-shift can produce low optical loses at second harmonics. The chromophores were successfully attached to a polyimide, yielding side chain polymers. The nonlinear optical property of the prepared optical polymers was determined by measuring electro-optic coefficient at 1.55 mm. The polymers exhibited high glass transition temperature of over $185^{\circ}C$ and thermal stability to $300^{\circ}C$ through differential scanning calorimeter analysis and thermal gravimetric analysis.

• Journal of Microbiology
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• v.44 no.5
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• pp.556-561
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• 2006

A differential display for the expression profiles of wild-type Cryphonectria parasitica and its virally-infected isogenic hypovirulent strain revealed several transcripts of interest, which evidenced significant matches with fungal genes of known function. Among which, we have further analyzed an amplified PCR product with significant sequence similarity to the known fungal stress-responsive thioredoxin gene from Neurospora crassa. The product of the cloned thioredoxin gene, CpTrx1, consists of 117 amino acids, with a predicted molecular mass of 13.0 kDa and a pI of 5.4. Sequence comparisons demonstrated that the deduced protein sequence of the CpTrx1 gene evidenced a high degree of homology to all known thioredoxins, with the highest degree of homology with trx1, a thioredoxin gene from Saccharomyces cerevisiae, and evidenced a preservation of the conserved hall markresidues (Trp-Cys-Gly-Pro-Cys) at the active site of thioredoxin. The E. coli-generated CpTRX1 manifested thioredoxin activity, according to the insulin reduction assay, which indicates that the cloned gene does indeed encode for the C. parasitica thioredoxin.

• Animal cells and systems
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• v.14 no.1
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• pp.25-30
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• 2010

We employed a new and improved differential display reverse transcription-polymerase chain reaction (DDRT-PCR) method, which involves annealing control primers (ACPs), to identify the genes that are specifically or prominently expressed in olive flounder (Paralichthys olivaceus) juveniles (35 days post-hatch; dph) compared to larval-stage (dph 21) flounder. Using 60 ACPs, we identified eight differentially expressed genes (DEGs) and basic local alignment search tool (BLAST) searches revealed eight known genes. Gene expression levels were confirmed by RT-PCR. Phosphoglucose isomerase (PGI) was highly expressed at 21 dph, while nephrosin, myosin light chain (MLC), myosin heavy chain (MHC), carboxypeptidase A, chymotrypsin B, fish-egg protein, and matrix protein were expressed at 35 dph. PGI, MLC, and MHC expression was further analyzed by RT-PCR. The differentially expressed genes identified in this study may provide insights into the molecular basis of development in olive flounder.

• Journal of Fisheries and Marine Sciences Education
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• v.29 no.3
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• pp.757-766
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• 2017

This study investigated not only the existence of differently functioned item due to gender but also domain. In this study, the randomly selected data of TIMSS 2007, which consist of 681 male and 646 women, were analyzed. To detect differently functioned items, this study employed Raju method. For Raju method, three-parameter logistic model was selected. Signed and unsigned area between two item characteristic curve were measured within the real ability range. An item which was detected commonly SA and UA area in Raju method was defined as a differently functioned item. As a result of this study, six items among twenty seven items of mathematics in the TIMSS 2007 were differently functioned item. Five items among those six items, were in favor of boys and one item was in favor of girls. Number, Geometric Shapes and Measures, and Applying were in favor of boys. but Data Display, Reasoning were in favor of girls. The conclusion of this study was summarized as existing differently functioned items in TIMSS 2007 and difference between favorable domain based gender. Finally, it is desirable to consider the differently functioned items by relating those item content for improving the test reliability of TIMSS 2007.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.74.1-74
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• 2003

Differential display (DD) of mRNA is a technique in which mRNA species expressed by a cell population are reverse transcribed and then amplified by many separate polymerase chain reactions (PCR). Using DD-RT-PCR we obtained many genes that expressed differentially in healthy and PVX-infected Nicotiana benthamima, using total RNAs extracted from healthy and PVX-infected N. benthamiana plants. Three hundred and twenty-five DNA fragments isolated from DD-RT-PCR were cloned and sequenced for further characterization. Several host genes including SKPI-like protein, heat shock transcription factor and Avr9/Cf-9 rapidly elicited protein were selected to obtain full-length open reading frame and to characterize their potential involvement in virus disease development and/or host's defense against virus infection employing PVX-based expression vector. Transcrips from wild-type and clones containing each selected gene were inoculated onto N. benthamiana Levels of virus replication were confirmedby RT-PCR and RNA blot analysis, Expression profiles and potential role(s) of selected genes upon PVX infection will be discussed.