• Journal of Plant Biotechnology
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• 제4권1호
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• pp.7-15
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• 2002

Eight cDNAs corresponding to fruit-specific genes were isolated from ripened melon through differential screening. Sequence comparison indicated that six of these cDNAs encoded proteins were previously characterized into aminocyclopropane-1-carboxylate (ACC) oxidase, abscisic acid, stress and ripening inducible (ASR) gene, RINC-H2 zinc finger protein, pyruvate decarboxylase, or polyubiquitin. RFS2 and RFS5 were the same clone encoding polyubiquitin. The other cDNAs showed no significant homology with known protein sequences. The ASR homologue (Asr1) gene was further characterized on the cDNA and genomic structure. The deduced amino acid sequence had similar characteristics to other plant ASR. The Asr1 genomic DNA consisted of 2 exons and 1 intron, which is similar to the structure of other plants ASR genes. The promoter region of the Asr1 gene contained several putative functional cis-elements such as an abscisic acid responsive element (ABRE), an ethylene responsive element (ERE), a C-box or DPBf-1 and 2, Myb binding sites, a low temperature responsive element (LTRE) and a metal responsive element (MRE). The findings imply that these elements may play important roles in the response to plant hormones and environmental stresses in the process of fruit development. The results of this study suggest that the expressions of fruit specific and ripening-related cDNAs are closely associated with the stress response.

• Molecules and Cells
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• 제39권2호
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• pp.149-155
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• 2016

In the heart, excitation-contraction (E-C) coupling is mediated by $Ca^{2+}$ release from sarcoplasmic reticulum (SR) through the interactions of proteins forming the $Ca^{2+}$ release unit (CRU). Among them, calsequestrin (CSQ) and histidine-rich $Ca^{2+}$ binding protein (HRC) are known to bind the charged luminal region of triadin (TRN) and thus directly or indirectly regulate ryanodine receptor 2 (RyR2) activity. However, the mechanisms of CSQ and HRC mediated regulation of RyR2 activity through TRN have remained unclear. We first examined the minimal KEKE motif of TRN involved in the interactions with CSQ2, HRC and RyR2 using TRN deletion mutants and in vitro binding assays. The results showed that CSQ2, HRC and RyR2 share the same KEKE motif region on the distal part of TRN (aa 202-231). Second, in vitro binding assays were conducted to examine the $Ca^{2+}$ dependence of protein-protein interactions (PPI). The results showed that TRN-HRC interaction had a bell-shaped $Ca^{2+}$ dependence, which peaked at pCa4, whereas TRN-CSQ2 or TRN-RyR2 interaction did not show such $Ca^{2+}$ dependence pattern. Third, competitive binding was conducted to examine whether CSQ2, HRC, or RyR2 affects the TRN-HRC or TRN-CSQ2 binding at pCa4. Among them, only CSQ2 or RyR2 competitively inhibited TRN-HRC binding, suggesting that HRC can confer functional refractoriness to CRU, which could be beneficial for reloading of $Ca^{2+}$ into SR at intermediate $Ca^{2+}$ concentrations.

• 한국잠사곤충학회지
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• 제40권2호
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• pp.111-116
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• 1998

To construct transformed Bm5 cells, Autographa californica nuclear polyhedrosis virus (AcNPV)IE1 gene, an immediate early viral gene was firstly used in this study. AcNPV IE1 gene, which shares on 95.3% uncleotide sequence homology with Bombyx mori nuclear polyhedrosis virus (BmNPV) IE1 gene, was isolated and cloned into pBluescript. Neomycin gene from pKO-neo was inserted under the control of the IE1 promoter to yield pAcIE1-neo. The plasmid pAcIE1-neo was transfected into Bm5 or Sf9 cells, and neomycin-resistant cells were selected in TC100 medium containing 10% fetal bovine serum (FBS) and 1 mg/$m\ell$ G418 for two weeks. Individual clones were picked and each was amplified for further characterization. The genomic DNA from neomycin-resistnt cells was isolated and characterized by PCR using AcNPV IE1 gene-specific primers and by Southern blot analysis using neomycin gene probe. We concluded that AcNPV IE1 gene was functional in B. moridrived Bm5 cells as well as Spodaptera frugiperda-derived Sf9 cells to produce stably-transformed insect cells.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2007년도 Proceedings of The Convention of The Korean Society of Applied Pharmacology
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• pp.147-153
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• 2007

Central to developing new treatment strategies for late onset sporadic Parkinson's disease (PD) and early onset familial PD is resolving the enigma of the specific vulnerability exhibited by substantia nigra dopamine (DA) neurons despite multiple risk factors. Neuropathological evidence from both human and experimental models of PD firmly supports a significant role for oxidative stress (OS) and mitochondrial dysfunction in the death of nigral DA neurons. Largely unknown are the genes underlying selective susceptibility of nigral DA neuron to OS and mitochondrial dysfunction and how they effect nigral DA cell death. To overcome the paucity of nigral DA neurons as well as the dilution effect of non-DA cells in brain tissues, we have developed wild type DA cell line model, SN4741 and mutant DJ-1 (-/-) DA cells, appropriate for microarray analysis and differential mitochondrial proteomics. Mutations in the DJ-1 gene (PARK7), localized in cytoplasm and mitochondria, cause autosomal recessive early onset PD. Through microarray analysis using SN4741 cells followed by validation tests, we have identified a novel phylogenically conserved neuroprotective gene, Oxi-a, which is specifically expressed in DA neurons. The knockdown of the gene dramatically increased vulnerability to as. Importantly as down-regulated the expression level of the gene and recovery of its expression via transient transfection exerted significant neuroprotection against as insult. We also have identified altered expression of mitochondrial proteins and other familial PD genes in DJ-1 (-/-) mutant cells by differential mitochondrial proteomics. In DJ-1 (-/-) cells the knockdown of the other familial PD genes (Parkin and PINK1) dramatically increased susceptibility to as. Thus, further functional characterization of the Oxi-$\alpha$ gene family and the mitochondrial alteration in the DJ-1 (-/-) cell model will provide the rationale for the neuroprotective therapy against both sporadic and familial PD.

• Journal of Plant Biotechnology
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• 제39권3호
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• pp.212-218
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• 2012

Glycosyltransferases are enzymes (EC 2.4) that catalyze the transfer of monosaccharide moieties from activated nucleotide sugar to a glycosyl acceptor molecule which can be a carbohydrate, glycoside, oligosaccharide, or a polysaccharide. In this study, a UDP-glucosyltransferase cDNA was isolated from Brassica rapa using a rapid amplification of cDNA ends (RACE) and subsequently named BrUGT. It has a full-length cDNA of 1,236 bp with 119 bp 5'-untranslated region (UTR), a complete ORF of 834 bp encoding a polypeptide of 277 amino acids (31.19 kDa) and a 3'-UTR of 283 bp. BLASTX analysis hits a catalytic domain of Glycos_transf_1 super family (cl12012) that belongs to the Glycosyltransferases group 1 with tetratricopeptide (TPR) regions located between 165 to 350 bp. Expression analysis showed high mRNA transcripts in pistil, followed by petal, seed and calyx of flower. Moreover, expression analysis of BrUGT in Chinese cabbage seedlings under stresses of cold, salt, PEG, $H_2O_2$, drought and ABA showed elevated mRNA transcript. Furthermore, when BrUGT gene was transformed into rice using pUbi-1 promoter, overexpression was evident among the $T_1$ plants. This study provides insights into the function of BrUGT in plants.

• 미생물학회지
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• 제35권1호
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• pp.28-34
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• 1999

본 연구에서는 Yeast Artificial Chromosome을 효율적으로 조작하고 유유전자의 기능을 보다 더 용이하게 분석하기 위해 EMCV 바이러스의 IRES 염기서열과 $\beta$-galactosidase를 포함하는 새로운 bicistronic fragmentation vector를 제조하였다. 이 벡터의 폴리클로닝 site에 네 개의희귀한 제한효소 부위를 도입하여 DNA를 용이하게 클로닝할 수 있게 만들었다. 그러므로 원하는 어떤 YAC도 효모세포에서 쉽게 상동 재조함에 의해 절편할 수 있는 장점이 있다. 이 bicistronic fragmentation vector 시스템을 이용하면하나의 메시지로부터 연구하고자 하는 유전자와 $\beta$-galactosidase를 동시에 한 세포에서 발현할 수 있기 때문에 유전자의 발현양상을 쉽게 분석할 수 있는 새로운 도구로 이용할 수 있다.

• 한국발생생물학회지:발생과생식
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• 제18권4호
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• pp.251-258
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• 2014

Serine-arginine-rich nuclear protein LUC7L plays an important role in the regulation of myogenesis in mice. In the present study, we isolated and characterized the Korean rose bitterling Rhodeus uyekii Luc7l cDNA, designated RuLuc7l. The RuLuc7l cDNA is 1,688 bp long and encodes a 364-amino-acid polypeptide containing serine/arginine-rich region at the C-terminus. The deduced RuLuc7l protein has high amino acid identity (71-97%) with those of other species including human. Phylogenetic analysis revealed that RuLUC7L clustered with fish LUC7L proteins. The expression of RuLuc7l mRNA was high in the brain, kidney, and stomach of Korean rose bitterling. Expression of the RuLuc7l mRNA was detected from 1 day post-fertilization (dpf) and moderately increased until 21 dpf during the early development. Further investigations are required to elucidate the functional role of RuLUC7L in myogenesis in R. uyekii.

• 공업화학
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• 제19권5호
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• pp.457-465
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• 2008

인간의 질병을 치료하기 위한 여러 가지 의료 시스템의 개발이 생명공학의 발전과 함께 많은 연구가 이루어지고 있다. 또한 약물이나 유전자와 같은 생리활성물질을 체내에 안전하게 전달할 수 있는 시스템의 개발과 함께 이루어지고 있다. 이러한 시스템에 있어서 가장 중요한 것은 생체적합성 및 생체분해성 그리고 무독성의 특성을 가진 생체의료용 고분자를 개발하는 것이다. 천연고분자물질인 키토산은 좋은 생체적합성과 생체활성의 특성을 가지고 있어서 생체의료용 재료로 심도 있게 고려되어지고 있다. 키토산의 물성은 키틴의 결정성 구조에 따라 다르게 설명되므로 키틴의 구조적 분석에 대한 연구가 생체재료로써의 응용을 위해서 선행되어야 한다. 이러한 관점에서 본 총설에서는 키틴의 결정성 구조 분석, 키토산의 일반적인 물성 그리고 생체의료용 재료로써 저분자량 수용성 키토산의 가능성을 소개하였다. 또한 다양한 기능성기를 이용한 저분자량 수용성 키토산의 화학적인 개질을 약물전달체로써의 가능성을 강조하고 생체이용율의 향상을 위해 수행하였다.

• 공업화학
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• 제23권1호
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• pp.100-104
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• 2012

알긴산 올리고당은 생체에 다양한 생리활성효과를 나타내는 기능성 식품의 소재나 고부가가치 재료로서 활용 가능성이 높아 여러 응용 범위 내에서 사용될 수 있다. 알긴산은 해조류의 주요 구성성분으로 주로 갈조류에 많이 존재한다. 해조류로부터 다양하게 활용이 가능한 알긴산 올리고당을 제조하기 위해 Erwinia tasmaniensis 균주가 생성하는 효소를 이용하여 알긴산을 효소적으로 분해하고자 하였다. 따라서 본 연구에서는 E. tasmaniensis의 최적 배양조건과 균주가 생성해내는 알긴산 분해 효소의 성질 및 특성을 알아보았다. 실험 결과, 이 균주의 최적 배양을 위한 배지 내 알긴산의 최적 농도는 1.0%, 최적 시간은 36 h이었으며, 알긴산 분해 효소의 활성은 알긴산이 1.0% 포함된 배지에서 72 h동안 배양했을 때 가장 높았다. 이 효소의 최적 pH는 6.0, 최적 온도는 $20^{\circ}C$로 확인되었다. 또한 효소의 활성은 $20^{\circ}C$에서 60 min까지 지속됨을 확인했다.

• Food Science and Biotechnology
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• 제16권4호
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• pp.509-514
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• 2007

In this study, we optimized the production of ${\gamma}-polyglutamic$ acid (PGA) in soybean milk cakes (SMC) fermented with Bacillus subtilis GT-D and B. subtilis KU-A, to be utilized as a functional food ingredient. PGA production was dependent upon the glutamate content, fermentation time, and type of Bacillus sp. The consistencies of the SMCs fermented by B. subtilis GT-D and B. subtilis KU-A were highest after 36 hr of fermentation, and then decreased gradually. The SMC fermented by B. subtilis KU-A had a higher consistency than the SMC fermented by B. subtilis GT-D. In the presence of 10% defatted soy flour (DFS), 5% glutamate in the SMC was efficiently converted into polyglutamic acid (PGA) for 24 hr, indicating a conversion yield above 96%, but its conversion then decreased with higher concentrations of glutamate. The soluble solid content (mucilage) of the SMC fermented with B. subtilis KU-A was 9.5%(w/w), and composed of 65.6% PGA (Mw 1,536 kDa) and some polysaccharides. However, the SMC fermented with B. subtilis GT-D had a mucilage content of 7.8%(w/w), and was composed of 66.4% PGA (Mw 1,409 kDa), 11.5% levan, and some polysaccharides. The viscoelastic values of the mucilage obtained using B. subtilis KU-A were much higher than those of mucilage obtained using B. subtilis GT-D. Also, the G'-value (elastic modulus) was higher than the G"-value (viscous modulus).