• Title/Summary/Keyword: nucleic acid

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Proteome characterization of the liquid cultured tetraploid roots in Platycodon grandiflorum

  • Ko, Jung-Hee;Kwon, Soo Jeong;Roy, Swapan Kumar;Cho, Seong-Woo;Kim, Hag Hyun;Boo, Hee Ock;Woo, Sun-Hee
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2017년도 9th Asian Crop Science Association conference
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    • pp.125-125
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    • 2017
  • The roots of Platycodon grandiflorum are commonly used for treating bronchitis, asthma, tuberculosis, diabetes, and other inflammatory diseases. Since the molecular mechanism underlying the roots of the plant is unclear. Therefore, the present study was conducted to profile proteins from liquid cultured tetraploid roots of Platycodon grandi orum fl using high throughput proteome approach. Two-dimensional gels stained with CBB, a total of 659 differentially expressed proteins were identified from the liquid medium cultured tetraploid roots of which 32 proteins spots (${\geq}1.5-fold$) were sorted for mass spectrometry analysis. Out of these 32 proteins, a total of 15 proteins were up-regulated such as Serine carboxypeptidase-like 27, Transcription factor bHLH150, 60 kDa jasmonate-induced protein, Cytosolic Fe-S cluster assembly factor NBP35, Regulatory associated protein of TOR 2 and a total of 17 proteins were down-regulated such as Protein G1-like2, Phenylalanine ammonia-lyase, Fructokinase-2, Trihelix transcription factor GT-3a, Guanine nucleotide-binding protein alpha-1 subunit. However, the frequency distribution of identified proteins was carried out within functional categories based on molecular functions, cellular components, and biological processes. Functional categorization revealed that the most of the identified proteins from the explants were mainly associated with the nucleic acid binding, oxidoreductase, transferase activity, protein binding and hydrolase activity. In addition, the proteomic feedback of tetraploid roots of P. grandiflorum may potentially be used to understand the characteristics of proteins and their functions.

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Double primary lung adenocarcinoma diagnosed by epidermal growth factor receptor mutation status

  • Kwon, Oh Jung;Lee, Min Hyeok;Kang, Sung Ju;Kim, Seul Gi;Jeong, In Beom;Jeong, Ji Yun;Cha, Eun Jung;Cho, Do Yeun;Kim, Young Jin;Son, Ji Woong
    • Journal of Yeungnam Medical Science
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    • 제34권2호
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    • pp.270-274
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    • 2017
  • A nodular density was detected on a chest radiograph taken from a 57-year-old Korean woman who was visiting a hospital for a routine check. Chest computed tomography revealed a 4.8 cm lobulated mass in the right lung and another focal nodular lesion in the left lung; biopsies of both lungs revealed adenocarcinoma. We conducted DNA sequencing and peptide nucleic acid clamping to investigate the potential double primary lung cancer. The results verified that the mass in the right lung had a mutation in the epidermal growth factor receptor, whereas the nodule in the left lung had a wild-type sequence, showing that these two were genetically different cancers from one another. Thus, we demonstrate that genetic testing is useful in determining double primary lung cancer, and we herein report on this case.

Isolation and Characterization of a New Methanobacterium formicicum KOR-1 from an Anaerobic Digester Using Pig Slurry

  • Battumur, Urantulkhuur;Yoon, Young-Man;Kim, Chang-Hyun
    • Asian-Australasian Journal of Animal Sciences
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    • 제29권4호
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    • pp.586-593
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    • 2016
  • A new methanogen was isolated from an anaerobic digester using pig slurry in South Korea. Only one strain, designated KOR-1, was characterized in detail. Cells of KOR-1 were straight or crooked rods, non-motile, 5 to $15{\mu}m$ long and $0.7{\mu}m$ wide. They stained Gram-positive and produced methane from $H_2+CO_2$ and formate. Strain KOR-1 grew optimally at $38^{\circ}C$. The optimum pH for growth was 7.0. The strain grew at 0.5% to 3.0% NaCl, with optimum growth at 2.5% NaCl. The G+C content of genomic DNA of strain KOR-1 was 41 mol%. The strain tolerated ampicillin, penicillin G, kanamycin and streptomycin but tetracycline inhibited cell growth. A large fragment of the 16S rRNA gene (~1,350 bp) was obtained from the isolate and sequenced. Comparison of 16S rRNA genes revealed that strain KOR-1 is related to Methanobacterium formicicum (98%, sequence similarity), Methanobacterium bryantii (95%) and Methanobacterium ivanovii (93%). Phylogenetic analysis of the deduced mcrA gene sequences confirmed the closest relative as based on mcrA gene sequence analysis was Methanobacterium formicicum strain (97% nucleic acid sequence identity). On the basis of physiological and phylogenetic characteristics, strain KOR-1 is proposed as a new strain within the genus Methanobacterium, Methanobacterium formicicum KOR-1.

Side Population Cell Level in Human Breast Cancer and Factors Related to Disease-free Survival

  • Jin, C.G.;Zou, T.N.;Li, J.;Chen, X.Q.;Liu, X.;Wang, Y.Y.;Wang, X.;Che, Y.H.;Wang, X.C.;Sriplung, Hutcha
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권3호
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    • pp.991-996
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    • 2015
  • Side population (SP) cells have stem cell-like properties with a capacity for self-renewal and are resistant to chemotherapy and radiotherapy. Therefore the presence of SP cells in human breast cancer probably has prognostic value. Objective: To investigate the characteristics of SP cells and identify the relationship between the SP cells levels and clinico-pathological parameters of the breast tumor and disease-free survival (DFS) in breast cancer patients. Materials and Methods: A total of 122 eligible breast cancer patients were consecutively recruited from January 1, 2006 to December 31, 2007 at Yunnan Tumor Hospital. All eligible subjects received conventional treatment and were followed up for seven years. Predictors of recurrence and/or metastasis and DFS were analyzed using Cox regression analysis. Human breast cancer cells were also obtained from fresh human breast cancer tissue and cultured by the nucleic acid dye Hoechst33342 with Verapami. Flow cytometry (FCM) was employed to isolate the cells of SP and non-SP types. Results: In this study, SP cells were identified using flow cytometric analysis with Hoechst 33342 dye efflux. Adjusted for age, tumor size, lymph nodal status, histological grade, the Cox model showed a higher risk of recurrence and/or metastasis positively associated with the SP cell level (1.75, 1.02-2.98), as well as with axillary lymph node metastasis (2.99, 1.76-5.09), pathology invasiveness type (1.7, 1.14-2.55), and tumor volume doubling time (TVDT) (1.54, 1.01-2.36). Conclusions: The SP cell level is independently associated with tumor progression and clinical outcome after controlling for other pathological factors. The axillary lymph node status, TVDT and the status of non-invasive or invasive tumor independently predict the prognosis of breast cancer.

3차원 배양 시스템을 이용한 돼지 유선 상피 세포 주 특성과 설정 (Establishment and characterization of porcine mammary gland epithelial cell line using three dimensional culture system)

  • 정학재
    • 한국산학기술학회논문지
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    • 제18권10호
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    • pp.551-558
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    • 2017
  • 조직특이적 프로모터 및 벡터를 연구하고 검증하기 위해서는 조직 및 종의 특이성을 유지하는 세포 시스템을 개발하는 것이 바람직하다. 이러한 시스템은 형질전환동물 모델에 대한 효과적인 대안이다. 우리는 베타 카제인 (CSN2)의 세포 형태와 mRNA 수준에 기초하여 일차 배양으로부터 돼지 유선 상피 세포 주 (PMECs)를 확립하였다. 선택된 PMECs는 cytokeratin 항체에 의해 염색되었으며, 유선 상피 세포에 존재한다고 생각되어지는 유즙 단백질 유전자 (CSN2, 락토페린 및 유청 단백질)를 발현하는 것으로 나타났다. 또한, 3D 배양에서 PMECs932-7의 acini 구조를 확인하기 위해 살아있는 세포를 핵산에 결합하는 SYTO-13으로 염색하였다. 우리는 마트리겔 (matrigel)에 있는 PMECs의 acini가 말초 세포의 응집에 의해 형성되고 공간의 lumen을 특징으로 한다는 것이 관찰하였다. 우리는 PMECs의 matrigel 사용법과 세포 밀도를 포함한 세포 배양 조건의 영향을 시험함으로서 시스템을 시연했다. 이러한 결과는 PMCEs의 유선 상피 세포는 유전적 또는 구조적 특징을 갖고 있음을 시사하고 있다.

숙시닐화가 Aspergillus fumiagtus 균체단백질의 기능적 특성에 미치는 영향 (Effect of Succinylation on Functional Properties of Aspergillus fumiagtus Cell Protein)

  • 최종덕;김정균;조성환
    • 한국식품영양과학회지
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    • 제21권5호
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    • pp.573-579
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    • 1992
  • 숙시닐화가 균체 단백질(Aspergillus fumigatus)의 여러가지 기능적 특성에 미치는 영향을 검토하였다. 균체 단백질은 succinic anhydride 2.5와 10% 첨가에 20.7 과 85.3%가 숙시닐화되었다. 숙시닐화된 균체 단백질은 흡광도, 핵산 및 탄수화물의 양은 감소하였으나 단백질소, 단백질 추출율은 증가되었다. 숙시닐화된 균체 단백질의 질소용해도는 milk casein과 soy flour 보다 32와 51% 증가되었다. 유화활성도와 유화 안정성은 균체 단백질의 숙시닐화 비율에 따라서 증가 되었으며, 80% 숙시닐화된 균체 단백질은 약 8.4배 증가율을 나타내어 숙시닐화를 증가시킴에 따라 유화활성도와 유화안정성이 증가 되었다. 숙시닐화 시킨 균체 단백질은 숙시닐화 시키지 않는 균체 단백질, milk casein 및 soy flour에 비하여 향상된 기능적 특성을 나타내었다.

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구강편평상피암종에서 상피성장인자 수용체의 과발현과 K-ras 유전자 변이 (Epidermal growth factor receptor overexpression and K-ras mutation detection in the oral squamous cell carcinoma)

  • 문병출;한세진;정동준;김경욱
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제37권5호
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    • pp.396-402
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    • 2011
  • Introduction: Epidermal growth factor is a single-chain polypeptide consisting of 53 amino acids with potent mitogenic activity that stimulates the proliferation of a range of normal and neoplastic cells through an interaction with its specific receptor (epidermal growth factor receptor, EGFR). This interaction plays a key role in tumor progression including the induction of tumor cell proliferation. An increased EGFR copy number have been associated with a favorable response to EGFR tyrosine kinase inhibitors therapy. In contrast, K-ras mutations tend to predict a poor response to such therapy. The aim of this study was to determine the correlation between the clinicopathological factors and the up-regulation of EGFR expression and Kras mutations in oral squamous cell carcinoma. Materials and Methods: This study examined the immunohistochemical staining of EGFR, K-ras mutation detection with peptide nucleic acid (PNA)-based real-time polymerase chain reaction (PCR) clamping in 20 specimens from 20 patients with oral squamous cell carcinoma. Results: 1. In the immunohistochemical study of poorly differentiated and invasive oral squamous cell carcinoma, a high level of EGFR staining was observed. The correlation between immunohistochemical EGFR expression and histological differentiation, as well as the tumor size of the specimens was significant (Pearson correlation analysis, significance [r] >0.5, P<0.05). 2. In PNA-based real-time PCR clamping analysis, a K-ras mutation was not detected in all specimens. Conclusion: These findings suggest that the up-regulation of the EGFR may play a role in the progression and invasion of oral squamous cell carcinoma that is, independent of a K-ras mutation.

식이제한이 흰쥐의 체내대사에 미치는 영향 (The Effect of 50% dietary restriction on Protein and Nucleic acid Metabolism of Albino rats)

  • 최길자;김숙희
    • Journal of Nutrition and Health
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    • 제3권3호
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    • pp.167-178
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    • 1970
  • 체중이 $43{\pm}2g$ 되는 흰쥐 암수 70마리를 7 group으로 나누어 Sugar-casein diet를 사료로 하여 pair-feeding에 의한 50% 식이제한으로 9주간 사육한 실험결과는 다음과 같다. 50%식이제한으로 나타나는 현상은 체중감소와 더불어 각 장기의 무게가 감소하였고 실험기간이 길어질수록 감소율이 높았다. 각 장기중 간과 비장의 무게의 감소가 심했고 뇌가 가장 영향을 적게 받았다. 질소의 체내 보유율은 식이제한 group이 control group 보다 높았으나 몸무게 g당 체내 보유율은 식이제한 group이 높았고 식이제한 group의 몸무게는 감소하여 신체 내에서 단백질의 경제적연 이용을 알 수 있다. 식이제한으로 인한 각장기의 질소 RNA, DNA함량은 비장이 식이제한에 가장 예민하고 뇌가 가장 둔한 경향을 나타내었다. 식이제한으로 Hematology에 뚜렷한 변화는 없고 식이제한으로 anemia의 현상이 나타난다고는 할 수 없고 Serum albumin 이나 A/G Ratio도 두 group 사이에 식이제한 전기간을 통해 차이가 없었다.

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Proteome Profiling Unfurl Differential Expressed Proteins from Various Explants in Platycodon Grandiflorum

  • Kim, Hye-Rim;Kwon, Soo-Jeong;Roy, Swapan Kumar;Cho, Seong-Woo;Kim, Hag-Hyun;Cho, Kab-Yeon;Boo, Hee-Ock;Woo, Sun-Hee
    • 한국작물학회지
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    • 제60권1호
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    • pp.97-106
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    • 2015
  • Platycodon grandiflorum, commonly known as Doraji in Korea, has a wide range of pharmacologic properties, such as reducing adiposity and hyperlipidemia, and antiatherosclerotic effects. However, the mechanisms underlying these effects remain unclear. In order to profile proteins from the nodal segment, callus, root and shoot, high throughput proteome approach was executed in the present study. Two dimensional gels stained with CBB, a total of 84 differential expressed proteins were confirmed out of 839 protein spots using image analysis by Progenesis SameSpot software. Out of total differential expressed spots, 58 differential expressed protein spots (${\geq}$ 2-fold) were analyzed using MASCOT search engine according to the similarity of sequences with previously characterized proteins along with the UniProt database. Out of 58 differential expressed protein, 32 protein spots were up-regulated such as ribulose-1,5-bisphosphate carboxylase, endoplasmic oxidoreductin-1, heat stress transcription factor A3, RNA pseudourine synthase 4, cysteine proteinase, GntR family transcriptional regulator, E3 xyloglucan 6-xylosyltransferase, while 26 differential protein spots were down-regulated such as L-ascorbate oxidase precursor, late embryogenesis abundant protein D-34, putative SCO1 protein, oxygen-evolving enhancer protein 3. However, frequency distribution of identified proteins using iProClass databases, and assignment by function based on gene ontology revealed that the identified proteins from the explants were mainly associated with the nucleic acid binding (17%), transferase activity (14%) and ion binding (12%). In that way, the exclusive protein profile may provide insight clues for better understanding the characteristics of proteins and metabolic activity in various explants of the economically important medicinal plant Platycodon grandiflorum.

해조류로부터 Arbitrary 및 ITS Primer들을 사용한 직접 PCR 유전자 증폭반응의 한계 (Limits of Direct PCR Amplification from Seaweeds Using Arbitrary and ITS Primers)

  • 김용국;진형주;박선미;진덕희;홍용기
    • 생명과학회지
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    • 제9권1호
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    • pp.15-21
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    • 1999
  • PCR 기법을 응용한 RAPD 방법은 대상 생물의 유전정보를 전혀 모르는 상태에도 arbitrary primer를 이용하여 증폭함으로써 생물 종간의 유전적 표식자를 확인할 수 있는 간편하고 유익한 유전분석 방법이다. 이같은 RAPD 방법을 이용하여 해조류 방사무늬 김, 미역, 구멍갈파래에 대하여 DNA를 추출하지 않고 직접 생 엽체 및 생 사상체, 생 배우체를 PCR template로 사용하여 PCR product를 생산하였다. 그러나 specific primer를 이용한 nulear r DNA의 internal trancribed spacer (ITS) 부위는 생성물을 만들지 못하였다. 간편한 LiCl 방법에 의하여 추출된 DNA를 사용하였을 때는 IST 및 RAPD 모두 PCR product를 생산하였다. 방사무늬 김의 엽체 (haploid)와 사상체 (dip-loid)로 부터의 ITS 생성물은 동일하였으나 RAPD 생성물은 사용한 arbitrary primer에 따라 36-50$\%$의 다른 band가 만들어졌다. 또한 직접 생 조직을 사용하였을때와 추출 DNA를 사용하였을 때도 53-57$\%$의 다른 band가 만들어 줬다. 그러므로 RAPD 방법으로 유전분석 실험에는 동일한 ploidy의 조직을 template로서 사용하는 것이 중요하다.

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