• Journal of Microbiology and Biotechnology
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• 제18권7호
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• pp.1235-1244
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• 2008

Indole-3-acetic acid (IAA) is produced commonly by plants and many bacteria, however, little is known about the genetic basis involving the key enzymes of IAA biosynthetic pathways from Bacillus spp. IAA intermediates from the Gram-positive spore-forming bacterium Paenibacillus polymyxa E681 were investigated, which showed the existence of only an indole-3-pyruvic acid (IPA) pathway for IAA biosynthesis from the bacterium. Four open reading frames (ORFs) encoding indole-3-pyruvate decarboxylase-like proteins and putative indole-3-pyruvate decarboxylase (IPDC), a key enzyme in the IPA synthetic pathway, were found on the genome sequence database of P. polymyxa and cloned in Escherichia coli DH5$\alpha$. One of the ORFs, PP2_01257, was assigned as probable indole-3-pyruvate decarboxylase. The ORF consisted of 1,743 nucleotides encoding 581 amino acids with a deduced molecular mass of 63,380 Da. Alignment studies of the deduced amino acid sequence of the ORF with known IPDC sequences revealed conservation of several amino acids in PP2_01257, essential for substrate and cofactor binding. Recombinant protein, gene product of the ORF PP2_01257 from P. polymyxa E681, was expressed in E. coli BL21 (DE3) as a glutathione S-transferase (GST)-fusion protein and purified to homogeneity using affinity chromatography. The molecular mass of the purified enzyme showed about 63 kDa, corresponding closely to the expected molecular mass of IPDC. The indole-3-pyruvate decarboxylase activity of the recombinant protein, detected by HPLC, using IPA substrate in the enzyme reaction confirmed the identity and functionality of the enzyme IPDC from the E681 strain.

• 한국균학회지
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• 제17권3호
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• pp.119-123
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• 1989

주름버섯목(目) 느타리버섯과 민주름버섯목(目) 잔나비걸상버섯과의 이목간(異目間) 원형질체(原形質體)를 polyethylene glycol로 유도하여 융합주(融合株) heterokaryon을 선발하였다. 버섯최소배지에서 극히 균사생장이 느렸으며 버섯완전배지에서 3번 계대배양되면서 다소 생장이 빨라졌다. 융합주 36균주의 75%는 양친의 균사가 혼합된 균총형태였으며 16.7%는 새로운 형태, 8.3%는 느타리버섯 형태이었다. 이들 중 양친의 균총(菌叢)이 혼합된 형태는 3번 계대배양 후 모두 느타리버섯 형태로 변하였다. 균사(菌絲)에는 클램프연결체가 없었고 원기(原基)도 형성하지 않았다. 융합주(融合株)를 전기영동법으로 esterase, malate dehydrogenase, peroxidase의 동위효소(同位酵素) 분석(分析)으로 비교하였는데 잔나비걸상버섯 효소는 뚜렷하지 않았으나 새로운 밴드의 형성으로 보아 두 양친 genome간의 상호작용이 존재하였다.

• Clinical and Experimental Reproductive Medicine
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• 제50권3호
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• pp.177-184
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• 2023

Objective: Reconstructed oocytes after polar body genome transfer constitute a potential therapeutic option for patients with a history of embryo fragmentation and advanced maternal age. However, the rescue of genetic material from the first polar body (PB1) through introduction into the donor cytoplasm is not yet ready for clinical application. Methods: Eighty-five oocytes were obtained following in vitro maturation (IVM) and divided into two groups: PB1 nuclear transfer (PB1NT; n=54) and control (n=31). Following enucleation and PB1 genomic transfer, PB1 fusion was assessed. Subsequently, all fused oocytes underwent intracytoplasmic sperm injection (ICSI) and were cultured in an incubator under a time-lapse monitoring system to evaluate fertilization, embryonic morphokinetic parameters, and cleavage patterns. Results: Following enucleation and fusion, 77.14% of oocytes survived, and 92.59% of polar bodies (PBs) fused. However, the normal fertilization rate was lower in the PB1NT group than in the control group (56.41% vs. 92%, p=0.002). No significant differences were observed in embryo kinetics between the groups, but a significant difference was detected in embryo developmental arrest after the four-cell stage, along with abnormal cleavage division in the PB1NT group. This was followed by significant between-group differences in the implantation potential rate and euploidy status. Most embryos in the PB1NT group had at least one abnormal cleavage division (93.3%, p=0.001). Conclusion: Fresh PB1NT oocytes successfully produced normal zygotes following PB fusion and ICSI in IVM oocytes. However, this was accompanied by low efficiency in developing into cleavage embryos, along with an increase in abnormal cleavage patterns.

• Genomics & Informatics
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• 제20권4호
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• pp.45.1-45.7
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• 2022

Food security will be affected by climate change worldwide, particularly in the developing world, where the most important food products originate from plants. Plants are often exposed to environmental stresses that may affect their growth, development, yield, and food quality. Auxin is a hormone that plays a critical role in improving plants' tolerance of environmental conditions. Auxin controls the expression of many stress-responsive genes in plants by interacting with specific cis-regulatory elements called auxin-responsive elements (AuxREs). In this work, we performed an in silico prediction of AuxREs in promoters of five auxin-responsive genes in Zea mays. We applied a data fusion approach based on the combined use of Dempster-Shafer evidence theory and fuzzy sets. Auxin has a direct impact on cell membrane proteins. The short-term auxin response may be represented by the regulation of transmembrane gene expression. The detection of an AuxRE in the promoter of prolyl oligopeptidase (POP) in Z. mays and the 3-fold overexpression of this gene under auxin treatment for 30 min indicated the role of POP in maize auxin response. POP is regulated by auxin to perform stress adaptation. In addition, the detection of two AuxRE TGTCTC motifs in the upstream sequence of the bx1 gene suggests that bx1 can be regulated by auxin. Auxin may also be involved in the regulation of dehydration-responsive element-binding and some members of the protein kinase superfamily.

• 대한임상검사과학회지
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• 제41권1호
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• pp.11-23
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• 2009

Chromosomal rearrangements are major pathology in hematological malignancies. The detection of minimal residual disease (MRD) for these gene rearrangements helps in monitoring treatment outcomes and predicting prognosis of patients. Recently, quantification of these gene transcripts based on real-time quantitative polymerase chain reaction (RQ-PCR) has been used as MRD detection. The purpose of this study is to ensure the usefulness of the RQ-PCR technique for detecting MRD in hamatological malignancy patients. The patients had been diagnosed to AML1-ETO positive AML, PML-RARa positive AML and BCR-ABL positive MPN at Chonnam National University Hwasun Hospital from Jan. 2006 to Aug. 2008. The fusion transcript was quntified by RQ-PCR and analyzed in comparison to conventional cytogenetics, FISH and RT-PCR. The fusion gene transcript was quantified by RQ-PCR in 57 samples from 14 patients with AML1-ETO positive AML, 79 samples from 27 patients with PML-RARa positive AML and 108 samples from 36 patients with CML. At diagnosis, the quantitative fusion transcripts for AM1-ETO, PML-RARa and BCR-ABL showed the range of 0.485552651~10.82233683 (mean 3.782217131, SD 2.998052348), 0.005300395~0.29267494 (mean 0.056901315, SD 0.080131381) and 0.1293929~12.94826849 (mean 1.701935665, SD 2.200913158). The increase of AML1-ETO fusion gene transcripts preceded morphologic relapse in two patients. Quantification of fusion gene transcripts by RQ-PCR could detected MRD in samples which were negative by in cytogenetic analysis or FISH. Our findings indicated that quantitative analysis of AML1-ETO, PML-RARa and BCR-ABL transcripts by RQ-PCR might be a useful tool for the monitoring of minimal residual disease in hematological malignancies.

• 한국균학회지
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• 제20권2호
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• pp.118-125
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• 1992

노랑느타리 Pleuroltus cornucopiae와 사철느타리 Pleurolus florida의 영양요구주(營養要求株)로부터 나출(裸出)한 원형질체(原形質體)를 polyethylene glycol로 융합(融合)하여 종간(種間) 체세포잡종(體細胞雜種)을 얻었다. 이들은 핵융합주(核融合株) nuclear hybrid 또는 이질이배체(異質二倍體) allodiploid, 이핵체(異核體) heterokaryon, 자연적(自然的) 분리성이핵체(分離性異核體) spontaneous segregated heterokaryon로 나누어졌으며, 6융합조합(融合組合)중에서 1조합(組合)에서만 꺽쇠연결체 clamp connection를 가진 것이 나타났다. 총 59 융합주 중에서 꺽쇠연결체를 가진 2균주와 가지지 않은 10균주가 완전한 자실체(子實體)를 형성하였는데 거의 사철느타리 형태(形態)와 유사(有似)하였으나 버섯의 갓 색택(色澤)이 양친(兩親)인 노랑느타리가 어릴때 yellow, 사철느타리가 orange white 인데비해 체세포 잡종은 light blond-greyish brown 으로 다양한 혼합색(混合色)으로 나타났다. 꺽쇠연결체가 없는 융합주로부터 발용(發茸)된 자실체는 두균주를 제외하고 모두 꺽쇠연결체를 가졌으며, 이 자실체(子實體) 조직(組織)을 완전배지(完全培地)에 배양(培養)한 결과 균사생장(菌絲生長)이 빠르고 균사도 꺽쇠연결체를 가져 본래(本來)의 융합주와는 다른 형태로 변하였다.

• Journal of Microbiology and Biotechnology
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• 제11권1호
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• pp.164-166
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• 2001

To examine the characteristics of the recombinant thin aggregative fimbriae of Salmonella, the AgfA subunit gene was amplified from Salmonella enteritidis using a PCR. The maltose binding protein (MBP)-AgfA fusion protein was overproduced in E. coli and purified. The secondary structure of AgfA was then elucidated from the difference CD spectra. An estimation of the secondary structure of AgfA using the self-consistent method revealed a mostly ${\beta}-sheet$ structure.

• 기술혁신학회지
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• 제6권4호
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• pp.508-517
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• 2003

Biochip, that is a fusion of BT/IT/NT, is the fundamental technology of bio-engineering in Post-genome Era. It is not only the tool to analyze the huge bio-informations but also the technology necessary for screening of new medicine and medical examination. Except some technological fields, biochip area has a good chance of occupying advantageous position in the world technology community by early aggressive investment, because a new access to this technology is possible without a past imitation process. This study is to analyze the trend of development of domestic and foreign biochip technology and its current market status. Finally it presents some suggestions for the government policy considering the possibility of its development in Korea.

• Journal of Microbiology and Biotechnology
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• 제23권7호
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• pp.923-931
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• 2013

Rapamycin, known as an inhibitor of Target of Rapamycin (TOR), is an immunosuppressant drug used to prevent rejection in organ transplantation. Despite the close association of the TOR signaling cascade with various scopes of metabolism, it has not yet been thoroughly investigated at the metabolome level. In our current study, we applied mass spectrometric analysis for profiling primary metabolism in order to capture the responsive dynamics of the Chlamydomonas metabolome to the inhibition of TOR by rapamycin. Accordingly, we identified the impact of the rapamycin treatment at the level of metabolomic phenotypes that were clearly distinguished by multivariate statistical analysis. Pathway analysis pinpointed that inactivation of the TCA cycle was accompanied by the inhibition of cellular growth. Relative to the constant suppression of the TCA cycle, most amino acids were significantly increased in a time-dependent manner by longer exposure to rapamycin treatment, after an initial down-regulation at the early stage of exposure. Finally, we explored the isolation of the responsive metabolic factors into the rapamycin treatment and the culture duration, respectively.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.20-20
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• 2017

Plant genomes include heterochromatic loci that consist of repetitive sequences and transposable elements. LTR retrotransposon is the major class of transposons in advanced plants in terms of proportion in plant genome. The elements contribute not only to genome size but also to genome stability and gene expression. A number of cases have been reported transposon insertions near genic regions affect crop traits such as fruit pigments, stress tolerance, and yields. Functional LTR retrotransposons produce extrachromosomal DNA from genomic RNA by reverse transcription that takes place within virus-like-particles (VLPs). DECREASED DNA METHYLATION 1 (DDM1) plays important roles in maintaining DNA methylation of heterochromatin affecting all sequence contexts, CG, CHG, and CHH. Previous studies showed that ddm1 mutant exhibits massive transcription of retrotransposons in Arabidopsis, but only few of them were able to create new insertions into the genome. RNA-dependent RNA POLYMERASE 6 (RDR6) is known to function in restricting accumulation of transposon RNA by processing the transcripts into 21-22 nt epigenetically activated small interfering RNA (easiRNA). We purified VLPs and sequence cDNA to identify functional LTR retrotransposons in Arabidopsis ddm1 and ddm1rdr6 plants. Over 20 LTR copia and gypsy families were detected in ddm1 and ddm1rdr6 sequencing libraries and most of them were not reported for mobility. In ddm1rdr6, short fragments of ATHILA gypsy elements were detected. It suggests easiRNAs might regulate reverse transcription steps. The highest enriched element among transposon loci was previously characterized EVADE element. It has been reported that active EVADE element is more efficiently silenced through female germline than male germline. By genetic analyses, we found ddm1 and rdr6 mutation affect maternal silencing of active EVADE elements. DDM1-GFP protein accumulated in megaspore mother cell but was not found in mature egg cell. The fusion protein was also found in early embryo and maternal DDM1-GFP allele was more dominantly expressed in the embryo. We observed localization of DDM1-GFP in Arabidopsis and DDM1-YFP in maize and found the proteins accumulated in dividing zone of root tips. Currently we are looking at cell cycle dependency of DDM1 expression using maize system. Among 10 AGO proteins in Arabidopsis, AGO9 is specifically expressed in egg cell and shoot meristematic cells. In addition, mutation of AGO9 and RDR6 caused failure in maternal silencing, implying 21-22 nt easiRNA pathway is important for retrotransposon silencing in female gametophyte or/and early embryo. On the other hand, canonical 24 nt sRNA-directed DNA methylation (RdDM) pathways did not contribute to maternal silencing as confirmed by this study. Heat-activated LTR retrotransposon, ONSEN, was not silenced by DDM1 but the silencing mechanisms require RdDM pathways in somatic cells. We will propose distinct mechanisms of LTR retrotransposons in germline and somatic stages.