• IMMUNE NETWORK
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• v.10 no.2
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• pp.35-45
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• 2010

Background: Expression of recombinant antibodies and their derivatives fused with other functional molecules such as alkaline phosphatase in Escherichia coli is important in the development of molecular diagnostic reagents for biomedical research. Methods: We investigated the possibility of applying a well-known Fos-Jun zipper to dimerize $V_H$ and $V_L$ fragments originated from the Fab clone (SP 112) that recognizes pyruvate dehydrogenase complex-E2 (PDC-E2), and demonstrated that the functional zipFv-112 and its alkaline phosphatase fusion molecules (zipFv-AP) can be produced in the cytoplasm of Origami(DE3) trxB gor mutant E. coli strain. Results: The zipFv-AP fusion molecules exhibited higher antigen-binding signals than the zipFv up to a 10-fold under the same experimental conditions. However, conformation of the zipFv-AP seemed to be influenced by the location of an AP domain at the C-terminus of $V_H$ or $V_L$ domain [zipFv-112(H-AP) or zipFv-112(L-AP)], and inclusion of an AraC DNA binding domain at the C-terminus of VH of the zipFv-112(L-AP), termed zipFv-112(H-AD/L-AP), was also beneficial. Cytoplasmic co-expression of disulfide-binding isomerase C (DsbC) helped proper folding of the zipFv-112(H-AD/L-AP) but not significantly. Conclusion: We believe that our zipFv constructs may serve as an excellent antibody format bi-functional antibody fragments that can be produced stably in the cytoplasm of E. coli.

• Molecules and Cells
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• v.23 no.1
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• pp.100-107
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• 2007

The moss Physcomitrella patens has two life cycles, filamentous protonema and leafy gametophore. A modified from of suppression subtractive hybridization (SSH), mirror orientation selection (MOS), was applied to screen genes differentially expressed in the P. patens protonema. Using reverse Northern blot analysis, differentially expressed clones were identified. The identified genes were involved mainly in metal binding and detoxification. One of these genes was an AP2 (APETALA2) domain-containing protein (PpACP1), which was highly up-regulated in the protonema. Alignment with other AP2/EREBPs (Ethylene Responsive Element Binding Proteins) revealed significant sequence homology of the deduced amino acid sequence in the AP2/EREBP DNA binding domain. Northern analysis under various stress conditions showed that PpACP1 was induced by ethephon, cadmium, copper, ABA, IAA, and cold. In addition, it was highly expressed in suspension-cultured protonema. We suggest that PpACP1 is involved in responses to metals, and that suspension culture enhance the expression of genes responding to metals.

• BMB Reports
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• v.43 no.8
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• pp.567-572
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• 2010

In this study, we cloned the ERF-B3 subfamily transcription factor gene BnaERF-B3-hy15 from Brassica napus L. Huyou15. This 600 bp gene encodes a 199 amino acid classic ethylene responsive factor (ERF), which shown no binding or very weak binding GCC box-binding activity by the yeast one-hybrid assay. We used gene shuffling and the yeast one-hybrid system to obtain three mutated sequences that can bind to the GCC box. Sequence analysis indicated that two residues, Gly156 in the AP2 domain and Phe62 at the N-terminal domain were mutated to arginine and serine, respectively. Changes of Gly156 to arginine and Phe62 to serine increased the GCC-binding activity of BnaERF-B3-hy15 and the alter of Gly156 to arginine changed the AP2-domain structure of BnaERF-B3-hy15.

• The Journal of Korean Physical Therapy
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• v.26 no.5
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• pp.351-357
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• 2014

Purpose: The purpose of this study was to investigate level of functioning in patients with stroke using Modified Bathel Index (MBI), World Health Organization Disability Assessment Schedule 2.0 (WHODAS 2.0), and ICF core-set for stroke. Methods: Sixty-four patients with stroke were recruited for this study from nine medical institutes. The ICF core-set for stroke, WHODAS 2.0, and MBI were used to collect subjects' functional levels. ICF core-set was employed here as a standard frame to observe multi-dimension of functioning, that is physiological bodily function, activity and participation (AP) in daily life, and current environmental factors (EF) in patients with stroke. WHODAS 2.0 and MBI were also used in order to have a specific functioning level for subjects. The linkage of each item in WHODAS 2.0 and MBI into the ICF core-set for stroke was examined. Pearson correlation coefficient was used for analysis of their relationships. Results: Functioning level of participants showed moderate resulting from MBI and WHODAS 2.0 ($73.48{\pm}22.27$ and $35.55{\pm}12.53$, respectively). Strong relationship was observed between ICF core-set and WHODAS 2.0, and with MBI. Each item of disability scales was obtained its linkage into ICF in the domain of AP. However, lack of correlation between MBI and ICF in the domain of EF was found due to absence of related factors. Conclusion: MBI was found to be linked mainly into ICF in the domain of AP and to have limited linkage into EF. Therefore, it should be suggested that the ICF concept frame should be used as a multi-dimensional approach to patients with stroke.

• Korean Journal of Microbiology
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• v.51 no.2
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• pp.115-125
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• 2015

A monoclonal antibody AP64 IgM binds to human acute nonlymphocytic leukemia (ANLL) cell line HL60 and also cross-reacts with the homologous antigen in a rat ANLL cell. This antibody mediated by complement, has leukemia a suppression effect. In this study, we generated a recombinant single-chain variable domain fragment (ScFv) which were derived from $V_H$ and $V_L$ cDNA of AP64 IgM-secreting hybridoma by RT-PCR. The two variable regions were joined with a single 15 amino acid linker $(G_4S)_3$. This recombinant ScFv was expressed as a single polypeptide chain from Escherichia coli BMH 71-18. The recombinant ScFv was purified by applying the periplasmic extract to $Ni^+$-NTA-agarose affinity column and detected with westernblot. The purified recombinant ScFv recognized a surface antigen (about 30 kDa) of HL60 cell line which is the same antigen detected by parental AP64 IgM. But the affinity of ScFv for a surface antigen of HL60 was lower than that of the parental AP64 IgM, which needs to be further improved. Overall, the recombinant ScFv specific to HL60 might be a useful bioreagent for either diagnostic or therapeutic purposes.

• Plant Breeding and Biotechnology
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• v.5 no.2
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• pp.123-133
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• 2017

AP2/EREBP gene family consists of transcription factor genes with a conserved AP2 DNA-binding domain and is involved in various biological processes. AP2/EREBP gene families were identified through genome-wide searches in five Cucurbitaceae species including cucumber, wild cucumber, melon, watermelon, and bitter gourd, which consisted of more than 100 genes in each of the five species. The gene families were further divided into five groups including four subfamilies (ERF, DREB, AP2 and RAV) and a soloist group. Among the subfamilies, DREB subfamily which is known to be related to abiotic stress response was more analyzed and a total of 25 genes were identified as Cucurbitaceae homologues of Arabidopsis CBF/DREB1 genes which are important for abiotic stress-response and tolerance. In silico expression profiling using RNA-Seq data revealed diverse expression patterns of cucumber AP2/EREBP genes. AP2/EREBP gene families identified in this study will be valuable for understanding the stress response mechanism as well as facilitating molecular breeding in Cucurbitaceae crops.

• IEMEK Journal of Embedded Systems and Applications
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• v.19 no.2
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• pp.73-81
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• 2024

Recently, mobile manipulators have been utilized in agriculture industry for weed removal and harvest automation. This paper proposes a domain adaptive fruit detection method for harvest automation, by utilizing OWL-ViT model which is an open-vocabulary object detection model. The vision-language model can detect objects based on text prompt, and therefore, it can be extended to detect objects of undefined categories. In the development of deep learning models for real-world problems, constructing a large-scale labeled dataset is a time-consuming task and heavily relies on human effort. To reduce the labor-intensive workload, we utilized a large-scale public dataset as a source domain data and employed a domain adaptation method. Adversarial learning was conducted between a domain discriminator and feature extractor to reduce the gap between the distribution of feature vectors from the source domain and our target domain data. We collected a target domain dataset in a real-like environment and conducted experiments to demonstrate the effectiveness of the proposed method. In experiments, the domain adaptation method improved the AP50 metric from 38.88% to 78.59% for detecting objects within the range of 2m, and we achieved 81.7% of manipulation success rate.

• BMB Reports
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• v.43 no.11
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• pp.738-743
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• 2010

The c-Jun $NH_2$-terminal kinase (JNK) signaling pathway participates in many physiological functions. In the current study we reported the cloning and characterization of five novel JNK2 transcript variants, which were designated as $JNK2\alpha3$, $JNK2\alpha4$, $JNK2\beta3$, $JNK2\gamma1$ and $JNK2\gamma2$, respectively. Among them, $JNK2\alpha4$ and $JNK2\gamma2$ are potential non-coding RNA because they contain pre-mature stop codons. Both $JNK2\alpha3$ and $JNK2\beta3$ contain an intact kinase domain, and both encode a protein product of 46 kDa, the same as those of $JNK2\alpha1$ and $JNK2\beta1$. $JNK2\gamma1$ contains a disrupted kinase domain and it showed a disable function. When over-expressed in mammalian cells, $JNK2\alpha3$ showed higher activity on AP-1 than that of $JNK2\beta3$ and $JNK2\gamma1$. Furthermore, $JNK2\alpha3$ and $JNK2\beta3$ showed different levels of substrate phosphorylation, although they both could promote the proliferation of 293T cells. Our results further demonstrate that JNK2 isoforms preferentially target different substrates and may regulate the expression of various target genes.

• BMB Reports
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• v.38 no.4
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• pp.440-446
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• 2005

A cDNA that was rapidly induced upon abscisic acid, cold, drought, mechanical wounding and to a lesser extent, by high salinity treatment, was isolated from Arabidopsis seedlings. It was classified as DREB subfamily member based on multiple sequence alignment and phylogenetic characterization. Since it encoded a protein with a typical ERF/AP2 DNA-binding domain and was closely related to the TINY gene, we named it TINY2. Gel retardation assay revealed that TINY2 was able to form a specific complex with the previously characterized DRE element while showed only residual affinity to the GCC box. When fused to the GAL4 DNA-binding domain, either full-length or its C-terminus functioned effectively as a trans-activator in the yeast one-hybrid assay while its N-terminus was completely inactive. Our data indicate that TINY2 could be a new member of the AP2/EREBP transcription factor family involved in activation of down-stream genes in response to environmental stress.

• Journal of The Institute of Information and Telecommunication Facilities Engineering
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• v.3 no.2
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• pp.42-50
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• 2004

This paper describes a new approach to Internet host mobility. We argue that local mobility, the performance of existing mobile host protocol can be significantly improved. It proposes Fast Moving Detection schemethat based on neighbor AP channel information and moving detection table. And, it composes Local Area Clustering Path (LACP) domain that collected in AP's channel information and MN interface information. It stored the roaming table to include channel information and moving detection. Those which use the proposal scheme will need to put LACP information into the beacon or probe frame. Each AP uses scheme to inform available channel information to MN. From the simulation result, we show that the proposed scheme is advantageous over the legacy schemes in terms of theburst blocking probability and the link utilization.