• Microbiology and Biotechnology Letters
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• v.19 no.6
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• pp.568-574
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• 1991

The cloned glyceraldehyde-3-phosphate dehydrogenase (GAP) gene of Saccharomyces cereviszae (Holland et al., 1983) has been characterized. Based on the communication, we have also cloned 2.1 kb CAP DNA fragment and modified this fragment as a portable promoter. Two yeast expression vectors, one is YCp type vector being maintained at low copy number (1 or 2) and the other is YEp type vector at high copy number, have been constructed with the GAP promoter and the PH05' gene as a reporter. Our plasrnids were introduc,ed into S. cerevisiae HY-1, which has been improved. The $Trp^+$ transformants expressed APase activity efficiently and showed high level of PH05' transcripts.

• The Korean Journal of Food And Nutrition
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• v.10 no.2
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• pp.268-271
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• 1997

Human mitochondrial aldehyde dehydrogenase(ALDH2) is mainly responsible for the oxidation of acetaldehyde generated during alcohol oxidation in vivo. To investigate the role of ALDH2 in alcohol metabolism, it was needed to get solubilized enzyme. The cDNA of ALDH2 is isolated from cDNA library and ligated to several expression vectors for E. coli. At almost expression system to be constructed, the broad expression band of ALDH2 was detected. But, the large part of the expressed protein consisted as inclusion body, the yield of solubilized enzyme was not more tan 5% of the total expressed amount. Recombinant ALDH2 was verified from the several expression systems.

• Journal of Sericultural and Entomological Science
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• v.44 no.2
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• pp.69-73
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• 2002

For the practical use of nuecin protein, we tried to overexpress nuecin using Bm5 insect cell and Bombyx mori. We inserted nuecin cDNA into pBm10po1-Xa vector derived from B. mori nuclear polyhedrosis virus (BmNPV), and expressed in Bm5 cells and B. mori respectively. SDS-PAGE and Northern blot analysis showed an expressed of the protein when baculovirus expression vector system (BEVS) was used. The amount of intracellular protein is abundant, but the amount of extracellular protein is poor. The results suggest that the biologically active nuecin protein produced by using BEVS is poor because incresed level of misfolded nuecin by the strong promoter, polyhedrin and p 10 of BEVS, decrease the level of free chaperons and foldases by binding with them.

• Microbiology and Biotechnology Letters
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• v.26 no.2
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• pp.161-166
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• 1998

To study the usefulness of Se301 cells, which is originated from Spodoptera exigua and has susceptibility to the Autographa californica NPV (AcNPV), as a host for the AcNPV-based expression vector system, we compared the characteristics of AcNPV in Se301 and Sf-21 cells. The symptom by viral infection was similar in both of cells, but the ratio of polyhedra released from the cell was higher in Se301 cells than in Sf-21 cells. The overall PIB productivity of AcNPV was similar in both cells but the size of polyhedra was larger in Se301 cells. While the polyhedrin expression efficiency was about 2.4 times higher in Se301 cells than in Sf-21 cells, the viral growth was higher in Sf-21 cells. These results suggested that Se301 cell is very useful in the AcNPV-based expression system as a host.

• Korean Journal Plant Pathology
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• v.11 no.2
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• pp.173-178
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• 1995

동물계에서 항바이러스와관련된 dsRNA 의존성 인산화 효소(PKR)의 유전자를 식물체에서 발현시킬 경우 PKR에 의한 단백질합성 및 식물바이러스의 증식조절 가능성에 대한 기초자료를 확보하기 위하여 사람에서 분리된 PKR cDNA를 Agrobacterium 방법에 의하여 연초식물체(Nicotiana tabacum cv. Xanthi-nc)로 형질전환시켰다. HindIII/PstI처리에 의해 얻어지는 약 1.8kb의 phPKR cDNA절편을 일련의 유전자 조작 방법을 통하여 식물발현벡터인 pBI121에 도입하여, p12168을 재조합하였다. 이를 A. tumefaciens LBA 4404에 형질전환시켜 연초식물체형질 전환에 이용하였다. 2mg/l BA와 0.5mg/l NAA가 포함되고 100$\mu\textrm{g}$/ml의 kanamycin이 첨가된 MS배지에서 shooting시킨 후 phytohormone이 첨가되지 않은 MS배지상에서 rooting을 시켜 형질전환 연초식물체를 얻었으며, 형질전환식물체는 정상식물체와 유사한 생육양상을 나타내었다. 형질전환식물체의 유전자도입은 hPKR cDNA의 전사부여는 RT-PCR 방법에 의하여 확인되었다.

• Journal of Life Science
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• v.34 no.7
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• pp.493-499
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• 2024

Antimicrobial peptides are antimicrobial substances inherent in animals and plants, with strong antibacterial activity even in small amounts and with various other functions such as antiviral and antioxidant actions. Plants can be grown with just water and sunlight, allowing for their mass production at low costs. However, transforming a chloroplast into one that produces antimicrobial peptides, rather than growing plants, increases the amount of protein expression and minimizes contamination of the ecosystem because gene transfer by pollen does not occur. In that context, using transgenic plant chloroplasts to produce recombinant proteins increases protein degradation and reduces the solubility of proteins. To solve this problem, we fused SUMO, a fusion protein, with a recombinant protein. We also used a 6xHis tag to purify the fusion protein. The antimicrobial peptide stomoxyn is an antibacterial substance found in stable flies. Stomoxyn has an α-helix structure and is amphiphilic, which allows it to dissolve bacterial cell membranes. In this study, we constructed a transformation vector to express stomoxyn in both plant chloroplasts and Escherichia coli and used this vector to confirm the expression of stomoxyn in E. coli. The expression of the protein was then confirmed in E. coli using a transformation vector. The expressed stomoxyn was purified by nickel column and SUMOase treatment, and its antibacterial activity was confirmed using an agar diffusion assay. The EGFP gene was used to ensure that the transformed vector was inserted into the chloroplast.

• Korean Journal of Microbiology
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• v.30 no.5
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• pp.410-414
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• 1992

Numerical identification was carried out for an isolate of Streptomyces strain producing the extracellular p-lactamase inhibitor. Fifty taxonomic unit characters were tested and the data were analyzed numerically using the TAXON program. The isolate was identified to the major cluster 5 of Streptomyces and it was best matched to Streptomyces omiyaensis which is a synonym of Streptomyces exfoliatus. Therefore, it was concluded that the isolate was identified to be a strain (SMF 19) of Streptomyces exjbliatus.

• Korean Journal of Poultry Science
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• v.50 no.4
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• pp.261-266
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• 2023

The skeletal muscles of livestock play a crucial role as protein sources for humans, and the consumption of poultry meat is steadily increasing worldwide. Numerous genes, including myogenic regulatory factors, are involved in myogenesis, and precise regulation of them is essential. In this study, genes specifically expressed in muscles were selected, and their promoters were cloned and analyzed. The analysis of gene expression in various tissues of animals revealed that many genes exhibited specific expression patterns in skeletal muscles, with TNNT3, TNNC2, and MYF6 genes showing similar patterns in poultry. The promoter regions of three genes were amplified by polymerase chain reaction to sizes of 1.2 kb, 1.03 kb, and 1.43 kb, respectively. These fragments were then inserted at the front of the enhanced green fluorescent protein gene in vectors. It was confirmed that the sequences of three promoters closely matched the chicken genome sequences. Upon introducing vectors with each promoter into QM7 quail muscle cells, all three promoters successfully induced the expression of the green fluorescent protein. The brightness of the green fluorescence in each promoter was approximately seven times dimmer compared to the control, CMV-IE promoter. It is predicted that more than 230 transcription factors can bind to each promoter, especially various transcription factors expressed in muscles, including myogenic regulatory factors such as MYF5, MYOD, and MYOG. These promoters can be valuable for studying gene expression in poultry muscle cells, and further research is needed to precisely investigate the regulatory region of gene expression in promoters.

• Korean journal of applied entomology
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• v.38 no.2
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• pp.145-149
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• 1999

To develop the baculovirus expression vector system (BEVS) adopting p10 gene promoter of Spodoptera exigua nucleopolyhedrovirus (SeNPV), we characterized the p10 gene of SeNPV. The nucleotide sequence of 545 bases including the coding region of p10 gene was determined. Compared with the previously reported SeNPV p10 gene (Zuidema et al., 1993), 4 bases were different in the 5' and 3' flanking region but no difference was found in the coding region. The p10 gene was located within Xho I 1.5 Kb, Sph 1 2.4 Kb and Cla I 4.0 Kb fragments by Southern hybridization analysis. Also, the Sph I 2.4 Kb and the Cla I 4.0 Kb fragments were cloned and their restriction enzyme maps were determined.

• KSBB Journal
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• v.22 no.1
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• pp.16-21
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• 2007

Although Energy demands of modern society increase rapidly, current energy would be exhausted shortly. Therefore development of bio-ethanol production process from cellulose containing materials was extremly demanded. Therefore development of highly functional cellulase is requisite for this purpose. In this study cellobio-hydrolase (CBH1) gene from Trichorderma reesei was used to increase cellulase activity by directed evolution and highly functional cellobio-hydrolase was obtained and characterized.