• Korean Journal of Microbiology
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• v.52 no.3
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• pp.249-253
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• 2016

The inky cap, Coprinellus congregatus, produces several laccase isozymes during its life cycle: both hyphal tip laccase and sclerotial laccase are involved in the fungal development. When this fungus was transferred to an acid liquid medium (pH 4.0-4.5), a new laccase was synthesized and secreted into the culture supernatant. In order to examine its regulation by external pH, green fluorescent protein gene was ligated at the downstream of the promoters having different lengths. These expression vectors having different promoter lengths were inserted into the fungal transformation vector, pBARGEM7-1. These expression vectors were introduced to the mating type a1 and a2 monokaryons, and the transformants were selected by the phosphinothricin resistance. Transformant a1 (a1TF) and transformant a2 (a2TF) were mated with each other to generate homozygotic dikaryon transformants. All these transformants were grown in neutral liquid medium for 5 days, and then the whole cell homogenates were transferred to the acidic liquid medium (pH 4.1). After 36 h incubation at $25^{\circ}C$, cells were harvested for the analysis of GFP expression. GFP expression was detected in the transformant having full-length promoter (2.0 kb), but other transformants having shorter length promoter (shorter than 1.29 kb) failed to show the fluorescence. Therefore, the acid-responsive element in the laccase promoter should be localized between -2.0 kb ~ -1.29 kb region.

• Journal of the Institute of Electronics Engineers of Korea CI
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• v.46 no.1
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• pp.112-120
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• 2009

Face membership authentication is to decide whether an incoming person is an enrolled member or not using face recognition, and basically belongs to two-class classification where support vector machine (SVM) has been successfully applied. The previous SVMs used for face membership authentication have been trained and tested using image feature vectors extracted from member face images of each class (enrolled class and unenrolled class). The SVM so trained using image feature vectors extracted from members in the training set may not achieve robust performance in the testing environments where configuration and size of each class can change dynamically due to member's joining or withdrawal as well as where testing face images have different illumination, pose, or facial expression from those in the training set. In this paper, we propose an effective class discriminating feature vector-based SVM for robust face membership authentication. The adopted features for training and testing the proposed SVM are chosen so as to reflect the capability of discriminating well between the enrolled class and the unenrolled class. Thus, the proposed SVM trained by the adopted class discriminating feature vectors is less affected by the change in membership and variations in illumination, pose, and facial expression of face images. Through experiments, it is shown that the face membership authentication method based on the proposed SVM performs better than the conventional SVM-based authentication methods and is relatively robust to the change in the enrolled class configuration.

• Journal of Microbiology and Biotechnology
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• v.8 no.6
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• pp.676-684
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• 1998

Baculovirus transfer and expression vectors with Hyphantria cunea nuclear polyhedrosis virus (HcNPV) were constructed. An initial transfer vector, pHcEV, constructed using HcNPV was previously reported (Park et al. 1993. J. Kor. Soc. Viral. 23: 141-151). Herein, the size of the vector was properly reduced, and a functionally perfect vector was constructed and named pHcEV-IV (6.7 kb). The vector has a 2.2-kb HcNPV DNA sequence in the 5'-flanking region of the vector's polyhedrin gene promoter. The 1.8-kb HcNPV DNA sequence, poly A signal sequence, T3 primer sequence, and 13 multicloning site sequences, in order, were ligated in front of the translation start codon of the polyhedrin gene. The cloning indicating marker lacZ gene was inserted into the pHcEV-IV, named pHcEV-IV-lacZ, and transferred into the wild-type virus. Recombinant expression virus, lacZ-HcNPV, was constructed by replacing the lacZ gene in the pHcEV-IV-lacZ with the polyhedrin gene of the wild-type virus. The recombinant virus was isolated from blue plaques that produce $\beta$-galactosidase without polyhedra. The lacZ gene insertion was confirmed by Southern hybridization analysis. The expression of the lacZ gene in Spodoptera frugiperda cells infected with the lacZ-HcNPV was examined by SDS-PAGE and colorimetric assay. One 116-kDa LacZ protein band appeared on the PAGE. The production rate of the $\beta$-galactosidase was approximately 50 international units (IU) per min per ml between 2 to 5 days postinfection (p.i.). The highest activity occurred at five days p.i. was 170 IU/min/$m\ell$. The enzyme activity first appeared about 20 h p.i. as measured by colorimetric assay.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.6
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• pp.2813-2818
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• 2012

Background: Considerable evidence suggests that metadherin (MTDH) is a potentially crucial mediator of tumor malignancy and an important therapeutic target for simultaneously enhancing chemotherapy efficacy and reducing metastasis risk. Inhibition of MTDH expression by RNA interference has been shown in several previous research, but silencing MTDH expression by microRNA (miRNA) interference in breast cancer has not been established. In the present study, we investigated the role of MTDH-miRNA in down-regulation of proliferation, motility and migration of breast carcinoma cells. Methods: Expression vectors of recombinant plasmids expressing artificial MTDH miRNA were constructed and transfected to knockdown MTDH expression in MDA-MB-231 breast cancer cells. Expression of MTDH mRNA and protein was detected by RT-PCR and Western blot, respectively. MTT assays were conducted to determine proliferation, and wound healing assays and transwell migration experiments for cell motility and migration. Results: Transfection of recombinant a plasmid of pcDNA-MTDH-miR-4 significantly suppressed the MTDH mRNA and protein levels more than 69% in MDA-MB-231 breast cancer cells. This knockdown significantly inhibited proliferation, motility and migration as compared with controls. Conclusions: MTDH-miRNA may play an important role in down-regulating proliferation, motility and migration in breast cancer cells, and should be considered as a potential small molecule inhibitor therapeutic targeting strategy for the future.

• Journal of the Institute of Electronics Engineers of Korea CI
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• v.42 no.1
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• pp.1-7
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• 2005

Cluster analysis of microarray data has been often used to find biologically relevant Broups of genes based on their expression levels. Since many functionally related genes tend to be co-expressed, by identifying groups of genes with similar expression profiles, the functionalities of unknown genes can be inferred from those of known genes in the same group. In this Paper we address a novel clustering approach, called seed clustering, and investigate its applicability for microarray data analysis. In the seed clustering method, seed genes are first extracted by computational analysis of their expression profiles and then clusters are generated by taking the seed genes as prototype vectors for target clusters. Since it has strong mathematical foundations, the seed clustering method produces the stable and consistent results in a systematic way. Also, our empirical results indicate that the automatically extracted seed genes are well representative of potential clusters hidden in the data, and that its performance is favorable compared to current approaches.

• Journal of the Korean Institute of Intelligent Systems
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• v.19 no.6
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• pp.821-827
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• 2009

Human emotion can be reflected by their facial expressions. So, it is one of good ways to understand people's emotions by recognizing their facial expressions. General recognition system of facial expressions had selected interesting points, and then only extracted features without analyzing physical meanings. They takes a long time to find interesting points, and it is hard to estimate accurate positions of these feature points. And in order to implement a recognition system of facial expressions on real-time embedded system, it is needed to simplify the algorithm and reduce the using resources. In this paper, we propose a real-time recognition algorithm of facial expressions that project the grid points on an expression space based on Gabor wavelet feature. Facial expression is simply described by feature vectors on the expression space, and is classified by an neural network with its resources dramatically reduced. The proposed system deals 5 expressions: anger, happiness, neutral, sadness, and surprise. In experiment, average execution time is 10.251 ms and recognition rate is measured as 87~93%.

• Journal of the Korean Institute of Intelligent Systems
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• v.19 no.1
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• pp.1-5
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• 2009

This paper suggests to apply mirror-reflected method based 2D emotion recognition database to 3D application. Also, it makes facial expression of fuzzy modeling using probability of emotion. Suggested facial expression function applies fuzzy theory to 3 basic movement for facial expressions. This method applies 3D application to feature vector for emotion recognition from 2D application using mirror-reflected multi-image. Thus, we can have model based on fuzzy nonlinear facial expression of a 2D model for a real model. We use average values about probability of 6 basic expressions such as happy, sad, disgust, angry, surprise and fear. Furthermore, dynimic facial expressions are made via fuzzy modelling. This paper compares and analyzes feature vectors of real model with 3D human-like avatar.

• Convergence Security Journal
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• v.19 no.1
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• pp.67-78
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• 2019

Most existing facial expression recognition methods use a uniform grid method that divides the entire facial image into uniform blocks when describing facial features. The problem of this method may include non-face backgrounds, which interferes with discrimination of facial expressions, and the feature of a face included in each block may vary depending on the position, size, and orientation of the face in the input image. In this paper, we propose a variable-size block method which determines the size and position of a block that best represents meaningful facial expression change. As a part of the effort, we propose the way to determine the optimal number, position and size of each block based on the facial feature points. For the evaluation of the proposed method, we generate the facial feature vectors using LDTP and construct a facial expression recognition system based on SVM. Experimental results show that the proposed method is superior to conventional uniform grid based method. Especially, it shows that the proposed method can adapt to the change of the input environment more effectively by showing relatively better performance than exiting methods in the images with large shape and orientation changes.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.18
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• pp.7825-7829
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• 2014

Objective: To investigate the effect of high expression of XAF1 in vivo or in vitro on lung cancer cell growth and apoptosis. Methods: 1. The A549 human lung cancer cell line was transfected with Ad5/F35 - XAF1, or Ad5/F35 - Null at the same multiplicity of infection (MOI); (hereinafter referred to as transient transfected cell strain); XAF1 gene mRNA and protein expression was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting respectively. 2. Methyl thiazolyl tetrazolium (MTT) and annexin V-FITC/PI double staining were used to detect cell proliferation and apoptosis before and after infection of Ad5/F35 - XAF1 with Western blotting for apoptosis related proteins, caspase 3, caspase - 8 and PARP. 3. After the XAF1 gene was transfected into lung cancer A549 cells by lentiviral vectors, and selected by screening with Blasticidin, reverse transcription polymerase chain reaction (RT-PCR) and Western blotting were applied to detect mRNA and protein expression, to establish a line with a stable high expression of XAF1 (hereinafter referred to as stable expression cell strain). Twenty nude mice were randomly divided into groups A and B, 10 in each group: A549/XAF1 stable expression cell strain was subcutaneously injected in group A, and A549/Ctrl stable cell line stable expression cell strain in group B (control group), to observe transplanted tumor growth in nude mice. Results: The mRNA and protein expression of XAF1 in A549 cells transfected by Ad5/F35 - XAF1 was significantly higher than in the control group. XAF1 mediated by adenovirus vector demonstrated a dose dependent inhibition of lung cancer cell proliferation and induction of apoptosis. This was accompanied by cleavage of caspase -3, -8, -9 and PARP, suggesting activation of intrinsic or extrinsic apoptotic pathways. A cell strain of lung cancer highly expressing XAF1 was established, and this demonstrated delayed tumor growth after transplantation in vivo. Conclusion: Adenovirus mediated XAF1 gene expression could inhibit proliferation and induce apoptosis in lung cancer cells in vitro; highly stable expression of XAF1 could also significantly inhibit the growth of transplanted tumors in nude mouse, with no obvious adverse reactions observed. Therefore, the XAF1 gene could become a new target for lung cancer treatment.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.5
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• pp.2045-2049
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• 2012

Objective: To investigate the effect of down-regulation of Sentrin/SUMO-specific protease 1 (SENP1) expression on the apoptosis of human Burkitt lymphoma cells (Daudi cells) and potential mechanisms. Methods: Short hairpin RNA (shRNA) targeting SENP1 was designed and synthesized and then cloned into a lentiviral vector. A lentiviral packaging plasmid was used to transfect Daudi cells (sh-SENP1-Daudi group). Daudi cells without transfection (Daudi group) and Daudi cells transfected with blank plasmid (sh-NC-Daudi group) served as control groups. Flow cytometry was performed to screen GFP positive cells and semiquantitative PCR and Western blot assays were employed to detect the inference efficiency. The morphology of cells was observed under a microscope before and after transfection. Fluorescence quantitative PCR and Western blot assays were conducted to measure the mRNA and protein expression of apoptosis related molecules (caspase-3, 8 and 9). After treatment with $COCl_2$ for 24 h, the mRNA and protein expression of hypoxia inducible factor -$1{\alpha}$ (HIF-$1{\alpha}$) was determined. Results: Sequencing showed the expression vectors of shRNA targeting SENP1 to be successfully constructed. Following screening of GFP positive cells by FCM, semiqualitative PCR showed the interference efficiency was $79.2{\pm}0.026%$. At 48 h after transfection, the Daudi cells became shrunken, had irregular edges and presented apoptotic bodies. Western blot assay revealed increase in expression of caspase-3, 8 and 9 with prolongation of transfection (P<0.05). Following hypoxia treatment, mRNA expression of HIF-$1{\alpha}$ remained unchanged in three groups (P>0.05) but the protein expression of HIF-$1{\alpha}$ markedly increased (P<0.05). However, in the sh-SENP1-Daudi group, the protein expression of HIF-$1{\alpha}$ remained unchanged Conclusion: SENP1-shRNA can efficiently inhibit SENP1 expression in Daudi cells. SENP1 inhibition may promote cell apoptosis. These findings suggest that SENP1 may serve as an important target in the gene therapy of Burkitts lymphoma.