• Journal of Microbiology
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• 제42권1호
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• pp.32-36
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• 2004

We have previously isolated three synthetic lethal mutants from Schizosaccharomyces pombe in order to identify mutations in the genes that are functionally linked to spmex67 with respect to mRNA export. A novel rsm1 gene was isolated by complementation of the growth defect in one of the synthetic lethal mutants, SLMex1. The rsml gene contains no introns and encodes a 296 amino-add-long protein with the RING finger domain, a C3HC4 in the N-terminal half. The Δrsm1 null mutant is viable, but it showed a slight poly(A)$\^$＋/ RNA accumulation in the nucleus. Also, the combination of Δrsm1 and Δspmex67 mutations confers synthetic lethality that is accompanied by the severe poly(A)$\^$＋/ RNA export defect. These results suggest that rsm1 is involved in mRNA export from the nucleus.

• Molecules and Cells
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• 제40권8호
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• pp.577-586
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• 2017

Phytocystatins (PhyCYSs) are plant-specific proteinaceous inhibitors that are implicated in protein turnover and stress responses. Here, we characterized a PhyCYS from Arabidopsis thaliana, which was designated AtCYS5. RT-qPCR analysis showed that the expression of AtCYS5 in germinating seeds was induced by heat stress (HS) and exogenous abscisic acid (ABA) treatment. Analysis of the expression of the ${\beta}-glucuronidase$ reporter gene under the control of the AtCYS5 promoter showed that AtCYS5 expression during seed germination was induced by HS and ABA. Constitutive overexpression of AtCYS5 driven by the cauliflower mosaic virus 35S promoter led to enhanced HS tolerance in transgenic Arabidopsis, which was characterized by higher fresh weight and root length compared to wild-type (WT) and knockout (cys5) plants grown under HS conditions. The HS tolerance of AtCYS5-overexpressing transgenic plants was associated with increased insensitivity to exogenous ABA during both seed germination and post-germination compared to WT and cys5. Although no HS elements were identified in the 5'-flanking region of AtCYS5, canonical ABA-responsive elements (ABREs) were detected. AtCYS5 was upregulated in ABAtreated protoplasts transiently co-expressing this gene and genes encoding bZIP ABRE-binding factors (ABFs and AREB3). In the absence of ABA, ABF1 and ABF3 directly bound to the ABREs in the AtCYS5 promoter, which activated the transcription of this gene in the presence of ABA. These results suggest that an ABA-dependent pathway plays a positive role in the HS-responsive expression of AtCYS5 during seed germination and post-germination growth.

• Journal of Microbiology
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• 제45권4호
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• pp.344-349
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• 2007

We have isolated previously three synthetic lethal mutants in Schizosaccharomyces pombe, which genetically interact with mex67, in order to identify the genes involved in mRNA export. A novel nup97 gene was isolated by complementation of the growth defect in one of the synthetic lethal mutants, SLMex3. The nup97 gene contains one intron and encodes an 851 amino-acid protein that is similar to nucleoporins, Nppl06p in S. pombe and Nic96p in Saccharomyces cerevisiae. The nup97 gene is essential for vegetative growth, and nup97 null mutant harboring pREP41X-Nup97 showed $poly(A)^+$ RNA export defect when expression of nup97 is repressed in the presence of thiamine. These results suggest that nup97 is involved in mRNA export from the nucleus to cytoplasm.

• Journal of Ginseng Research
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• 제14권2호
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• pp.207-209
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• 1990

The sequence of ginseng peptide gene was designed and synthesized by the solid phase phosphoramidiate method. Synthetic segments were isolated, pllrified and joined to the plasmid pUC19. E.icherichiu coli JM101 cells were transformed with above hybrid plasmids. AmpiciIBin resistant transformants were screened and identified by in situ colony hybridization and Southern blot techniques. Finally the gene sequencing was done by the Sanger dideoxy method using primer extension.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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• pp.292.2-293
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• 2003

In the gene therapy. viral gene delivery systems are limited in use because of several drawbacks like host immune reactions. Hence, non-viral gene delivery systems such as cationic polymers or synthetic gene carriers are being widely investigated to overcome the problems in the use of viral vectors. We synthesized a new conjugate of polyethyleniminet carrying galactose moieties as a targeting ligand for asialoglycoprotein (ASGP) receptors of hepatocytes. (omitted)

• Molecules and Cells
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• 제27권2호
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• pp.225-235
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• 2009

Antibody phage display provides a powerful and efficient tool for the discovery and development of monoclonal antibodies for therapeutic and other applications. Antibody clones from synthetic libraries with optimized design features have several distinct advantages that include high stability, high levels of expression, and ease of downstream optimization and engineering. In this study, a fully synthetic human scFv library with six diversified CDRs was constructed by polymerase chain reaction assembly of overlapping oligonucleotides. In order to maximize the functional diversity of the library, a ${\beta}$-lactamase selection strategy was employed in which the assembled scFv gene repertoire was fused to the 5'-end of the ${\beta}$-lactamase gene, and in-frame scFv clones were enriched by carbenicillin selection. A final library with an estimated total diversity of $7.6{\times}10^9$, greater than 70% functional diversity, and diversification of all six CDRs was obtained after insertion of fully randomized CDR-H3 sequences into this proofread repertoire. The performance of the library was validated using a number of target antigens, against which multiple unique scFv sequences with dissociation constants in the nanomolar range were isolated.

• 식물조직배양학회지
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• 제22권4호
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• pp.201-205
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• 1995

Nopaline synthase promoter의 upstream element region을 본딴 nos-RP 요소라고 불리워진 합성 oligomer는 nos wild type promotor의 5' end에서부터 - 101까지를 절단한 promoter의 upstream에 삽입하였다. Nos promoter의 활성은 nos promoter와 연결되어 있는 reporter gene인 Chlorarnphenicol과 $\beta$-glucuronidase유전 인자들이 발현되는 현상을 연구함으로써 측정하였다. 형질 전환된 유전인자를 가지고 있는 담배 식물에 대한 분석은 nos minimal promoter의 활성이 합성 nos-RP 요소가 삽입됨으로써 회복될 수 있음을 보여 주었다. 또한 Nos minimal promoter의 upstream에 nos-RP 요소의 삽입은 여러 가지 환경적인 요소들인 auxin, dithiothreitol, salicylic acid 그리고 methyl jasmonate에 의해서 활성이 증가됨을 보여 주었다.

• 한국식물병리학회지
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• 제10권3호
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• pp.157-162
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• 1994

Erwinia rhapontici causes soft-rot disease in a number of plants such as onion, garlic and hyacinth. There has been no report that E. rhapontici produces pectate lyase. Pel gene was cloned from genomic DNA of the parasitic soft-rot E. rhapontici polymerase chain reaction by using synthetic oligonulceotide primers designed from the pel 1 to E. carotovora. The recombinant plasmid pJE101 containing pectate lyase gene, when introduced into E. coli DH5$\alpha$, produced pectate lyase an macerated hyacinth tissue.

• Journal of Microbiology and Biotechnology
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• 제21권7호
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• pp.719-727
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• 2011

Botulism is a neuroparalytic disease caused by Clostridium botulinum, which produces seven (A-G) antigenically diverse neurotoxins (BoNTs). BoNTs are the most poisonous substances known to humans, with a median lethal dose ($LD_{50}$) of approximately 1 ng/kg of body weight. Owing to their extreme potency and lethality, they have the potential to be used as a bioterrorism agent. The mouse bioassay is the gold standard for the detection of botulinum neurotoxins; however, it requires at least 3-4 days for completion. Attempts have been made to develop an ELISA-based detection system, which is potentially an easier and more rapid method of botulinum neurotoxin detection. The present study was designed using a synthetic gene approach. The synthetic gene encoding the catalytic domain of BoNT serotype B from amino acids 1-450 was constructed with PCR overlapping primers (BoNT/B LC), cloned in a pQE30 UA vector, and expressed in an E. coli M15 host system. Recombinant protein production was optimized at 0.5 mM IPTG final concentration, 4 h post induction, resulting in a maximum yield of recombinant proteins. The immunogenic nature of the recombinant BoNT/B LC protein was evaluated by ELISA. Antibodies were raised in BALB/c mice using various adjuvants. A significant rise in antibody titer (p<0.05) was observed in the Alum group, followed by the Titermax Classic group, Freund's adjuvant, and the Titermax Gold group. These developed high-titer antibodies may prove useful for the detection of botulinum neurotoxins in food and clinical samples.

• 대한화장품학회지
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• 제36권1호
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• pp.33-39
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• 2010

최근 합성 염모제의 부작용들 때문에 인체에 무해한 천연재료를 이용한 염색에 대한 관심이 고조되어 있다. 본 연구에서는 농산 폐기물인 칡뿌리 추출물을 이용해 탈색 모발에 천연 염색을 함으로써 염색에 적절한 염색 시간, 염색시 온도와 매염과 매염제 처리 농도 변화 등에 따른 모발의 염색 정도를 색차계를 통해 조사하였다. 합성 염모제가 가진 화학 물질로 인한 모발 손상에 비해 칡뿌리 추출물을 이용한 모발 염색은 손상도를 낮춰주고 큐티클을 부드럽게 유지시키는 결과를 가져왔다. 칡 추출물만으로 염색한 모발이 가장 어두워짐을 관찰함으로써, 매염제는 염색의 보조 역할을 하는 성분임을 알 수 있었다. 본 연구의 결과를 종합하여 볼 때 칡뿌리 추출물은 합성 염모제에 비해 모발 손상도가 적으며 칡뿌리 추출물을 천연 모발 염모제의 주원료로써 활용가치가 있음을 제안하는 바이다.