• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2002년도 국제심포지엄
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• pp.107-107
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• 2002

ADAM은 metalloprotease/disintegrin domain을 가진 transmembrane glycoprotein 으로서 지금까지 30개 이상의 ADAM 및 10개 이상의 ADAMTS 단백질이 알려져 있다. 이들의 기능은 포유동물의 수정시 sperm-egg binding과 fusion, myoblast fusion, integrin과의 결합 등에 직접 관여하거나, TNF-alpha 등의 생체 신호전달물질이 세포로부터 분비될 때에 이들의 구조를 변화시켜 활성화시키는 효소작용, 그리고 dendritic cell differentiation 등에 관여하는 것으로 알려져있다. 그러나 자궁내막 조직에서의 유전자 및 단백질 발현 여부에 관해서는 거의 보고되어 있지 않고 있다. 먼저 RT-PCR 방법을 이용하여 자궁에서 mRNA의 양을 $\beta$-actin의 양에 대하여 상대적으로 측정한 결과, 임신 1일부터 5일까지는 ADAM-9, 10, 17, TS1의 mRNA 경우 거의 비슷하게 발현되었으며 ADAM-8과 15는 3일째, ADAM-12는 2일째와 4일째에 현저하게 증가하였다 임신 6일부터 8일까지의 자궁조직을 각각 embryo가 착상된 곳과 그렇지 않은 곳으로 나누어 조사한 결과 상대적으로 착상이 된 곳에서 mRNA가 많이 발현 되었으며 특히 ADAM-8, 12, 15의 mRNA의 경우 현저한 차이를 보였다. Immunoblotting 방법을 이용하여 임신 1일부터 5일까지의 단백질의 발현 양상을 조사한 결과 임신 ADAM-8은 3일 이후 감소되는 양상을 보였으며 ADAM-9, 15, TS1은 5일째에 증가하는 양상을 보였다. ADAM-10은 2일째 감소하다가 다시 증가하고 ADAM-12, 17은 3, 4일째 증가하다가 5일째 감소하는 상을 보였다. 한편 임신 6일부터 8일까지는 embryo가 착상된 곳이 그렇지 않은 곳보다 조사된 모든 ADAMs 단백질이 상대적으로 많이 발현되는 양상을 보였다. 이러한 결과로 미루어 ADAMs은 임신 초기 착상과정과 임신 단계에 따른 자궁의 조직 재구성에 중요한 역할을 할 것으로 생각된다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권10호
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• pp.4153-4158
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• 2014

Background: This study was aimed to establish a novel method to simultaneously detect expression of four genes, ribonucleotide reductase subunit M1(RRM1), X-ray repair cross-complementing gene 1 (XRCC1), thymidylate synthase (TS) and class III ${\beta}$-tubulin (TUBB3), and to assess their application in the clinic for prediction of response of non-small cell lung cancer (NSCLC) to chemoradiotherapy. Materials and Methods: We have designed four gene molecular beacon (MB) probes for multiplex quantitative real-time polymerase chain reactions to examine RRM1, XRCC1, TUBB3 and TS mRNA expression in paraffin-embedded specimens from 50 patients with advanced or metastatic carcinomas. Twenty one NSCLC patients receiving cisplatin-based first-line treatment were analyzed. Results: These molecular beacon probes could specially bind to their target genes in homogeneous solutions. Patients with low RRM1 and XRCC1 mRNA levels were found to have apparently higher response rates to chemoradiotherapy compared with those with high levels of RRM1 and XRCC1 expression (p<0.05). The TS gene expression level was not significantly associated with chemotherapy response (p>0.05). Conclusions: A method of simultaneously detecting four molecular markers was successfully established and applied for evaluation of chemoradiotherapy response. It may be a useful tool in personalized cancer therapy.

• Molecules and Cells
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• 제42권5호
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• pp.379-385
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• 2019

Non-coding RNAs (ncRNAs) comprise various RNA species, including small ncRNAs and long ncRNAs (lncRNAs). ncRNAs regulate various cellular processes, including transcription and translation of target messenger RNAs. Recent studies also indicate that ncRNAs affect organismal aging and conversely aging influences ncRNA levels. In this review, we discuss our current understanding of the roles of ncRNAs in aging and longevity, focusing on recent advances using the roundworm Caenorhabditis elegans. Expression of various ncRNAs, including microRNA (miRNA), tRNA-derived small RNA (tsRNA), ribosomal RNA (rRNA), PIWI-interacting RNA (piRNA), circular RNA (circRNA), and lncRNA, is altered during aging in C. elegans. Genetic modulation of specific ncRNAs affects longevity and aging rates by modulating established aging-regulating protein factors. Because many aging-regulating mechanisms in C. elegans are evolutionarily conserved, these studies will provide key information regarding how ncRNAs modulate aging and lifespan in complex organisms, including mammals.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권3호
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• pp.273-276
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• 2006

RNA interference (RNAi), a process of sequence-specific gene suppression, has been known as a natural gene regulatory mechanism in a wide range of lower organisms. Recently, we have reported that a transfection of human U6 promoter (hU6) driven hairpin small-interference RNA (siRNA) plasmid specifically knocks down the target gene by post-transcriptional gene silencing in mammalian cells. Here we report that transfection of polymerase chain reaction (PCR) products, containing human U6 promoter with hairpin siRNA, knocks down the target gene expression in human teratocarcinoma NT-2/D1 cells. Moreover, we showed 3' end termination sequence, 5 Ts, is not critical elements for knocking down in PCR-based siRNA system. Therefore, the PCR-based siRNA system is a promising tool not only for the screening but also to temporally regulate gene expression in the human progenitor cells.

• Journal of Microbiology
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• 제35권4호
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• pp.334-340
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• 1997

A collection of 132 temperature sensitive (ts) mutants was generated by the chemical mutagenesis of Schizosaccharomyces pombe wild type strain and screened for tRNA splicing defects on Northern blots by hybridization with an oligonucleotide that recognizes the exon of the S. pombe tRNA^Val as a probe. We identidied 6 mutants which accumulate precursor $tRNA^{Val}$. Among them, 2 mutants exhibited remarkable morphological differences compared to wild type cells. One tRNA splicing mutant showed elongated cell shape in permissive as well as non-permissive cultures. The other mutant exhibited shortened cell morphology only in nonpermissive culture. The total RNA pattern in the splicing mutants appeared to be normal. Genetic analysis of four $tRNA^{Val}$ splicing mutants demonstrated that the mutation reside in different genes.

• Asian-Australasian Journal of Animal Sciences
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• 제25권5호
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• pp.648-652
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• 2012

This study was conducted to determine the effects of dietary addition of tea saponins (TS) and soybean oil (SO) on fatty acid profile and gene expression of stearoyl-CoA desaturase (SCD) in longissimus dorsi (LD) muscle of growing lambs. Thirty-two Huzhou lambs were assigned to four dietary treatments in a $2{\times}2$ factorial arrangement with main effects of TS (0 or 3 g/d) and SO (0 or 30 g/kg of diet DM). The diet without additives was considered as NTNS (no TS or SO). After a feeding trial for 60 d, four lambs of each treatment were slaughtered to collect the samples of LD muscle. Percentage of trans-11 vaccenic acid was enhanced (p<0.05) in muscle of lambs fed TS and SO. The proportion of total conjugated linoleic acid (CLA) was increased (p<0.05) by SO, but decreased (p<0.05) by TS in LD muscle. The percentage of total saturated fatty acids in muscle was decreased (p<0.05) by addition of TS and SO, while addition of SO increased (p<0.05) the percentage of total polyunsaturated fatty acids. The ratio of cis-9, trans-11 CLA to tran-11 vaccenic acid was decreased (p<0.05) by TS, but increased (p<0.05) by SO. The same effects were observed in SCD mRNA expression. From these results it is indicated that including TS and SO in the diet of growing lambs affect the fatty acid profiles of LD muscle and that the proportion of cis-9, trans-11 CLA in the muscle influenced by TS and SO may be related to the SCD gene expression.

• 미생물학회지
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• 제27권2호
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• pp.77-84
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• 1989

Saccharomyces cerevisiae의 RNAI 유전자의 온도 감수성 돌연변이 균주는 성장허용 온도인 23"C에서는 정상적인 성"J--을하나, 성 장억제 온도인 36°C에서는 tRNA, rRNA 그러고 mRNA 의 선우울질을을 핵내에 축적함으후써 성장을 못한다. 본 실 험에서는 complementation 에 의하여 RNAI 유전자를 클로녕하였으며 concomitant loss 실험에 의하여 이 유천자의 클로닝 을 확인하였다. 유전자의 위치를 확인한 결과 3.5kb의 Bgl II 조각내에 RNAI 유전자가 존재함을 알 수 있였으며, 2.1kb에 해당하는 BamH I -Bgl II 조각에서는 RNAI 유전지에 의한 complementation 능력이 상실되는 것으후 보아 RNAI 유전자 는 3.5kb의 BglIl 조각내에 포함되는 BamH 1 자리 주위에 결쳐 있픔을 알 수 있었다.

• 미생물학회지
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• 제27권3호
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• pp.176-180
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• 1989

E. coli의 $tRNA^{phe}$내에는 세 개의 psiudouridine 염기들이 존재한다. 이 $tRNA^{phe}$내의 pseudouridine 염기들의 기능을 연구하기 위하여 부위특이적 돌연변이를 이용하여 $tRNA^{phe}$ 유전자의 염기를 다른 염기로 치환시켰다. E. coli $tRNA^{phe}$ 유전자들 중 하나인 phe W 유전자내에서 32번에 해당하는 T 염기를 C 염기로 39번 T 염기를 C 염기로 Kunkel이 개발한 부위특이적 돌연변이 방법을 사용하여 각각 치환시켰다. DNA 염기서열을 결정함으로써 돌연변이체를 확인하였으며, 이들 돌연변이 유전자를 함유한 재조합 플라스미드를 이용하여 돌연변이된 phe W 유전자들의 E. Coli NP37($pheS^{-ts}$)에 대한 complementation 활성을 조사하였다. 32번 위치가 변이된 pheW 유전자 뿐만아니라 39번 위치가 변이된 phe W 유전자를 함유한 E. coh NP37들은 모두 non-permissive temperature에서 자라지 못하였다. 이 결과는 변이된 pheW 유전자들이 E. coli NP37을 complementation 할 수 없으며, 또 pseudouridine 염기들이 생체내에서 E. coli $tRNA^{phe}$의 활성에 필수적이라는 것을 의미한다.

• 한국수산과학회지
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• 제42권5호
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• pp.461-467
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• 2009

A bacterial strain was screened and identified from sea sediments in Tongyeong coastal area in order to obtain proteases or peptidase cleaving C-terminal of glutamic acid. Strain TS0611, which showed the highest activity from 5 isolated protease producing strains, was selected and its properties investigated. Strain TS0611 was a gram-positive rod, $1.2\;{\mu}m$ in cell length, catalase positive, motility-positive, melanin-negative and grew at 15~$50^{\circ}C$, and hydrolyzed gelatin and casein. This strain was identified as Bacillus thuringiensis or Bacillus cereus based on results from the API system(API 50 CHB) which examined saccharides properties, fatty acid composition of cell wall, and 16S rRNA gene sequence.

• 미생물학회지
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• 제29권2호
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• pp.75-79
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• 1991

The yeast L-A virus (4.6 kb dsRNA genome) encodes the major coat protein and a "gag-pol" fusion minor coat protein that separately encapsidate itself and $M_{1}$, a 1.8 kb dsRNA satellite virus encoding a secreted protein toxin (the killer toxin). The teast chromosomal SKI genes prevent viral cytopathology by lowering the virus copy number. Thus, $ski^{-}$ mutants are ts and cs for growth. We transformed a ski2-2 virus-infested mutant with a yeast bank in a high copy cloning vector and selected the rare healthy transformants for analysis. One type of transformant segregated M-O L-A-O cells with high frequency. Elimination of the DNA clone from the ski2-2 strain eliminated this phinotype and introduction of the DNA clone recovered from such transformants into the parent ski2-2 strain, or into ski3 or ski6 mutants gave the same phenotype. This killer-curing phenotype was due to the curing of the helper L-A dsRNA virus. The 6.5 kb insert only had this activity when carried on a high copy vector and in $ski^{-}$ cells (not in $SKI^{+}$ cells). This 6.5 kb insert acts as a mutagen on L-A dsRNA producing a high rate of deletion mutations.mutations.