• Title/Summary/Keyword: 단백질 발현

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Expression and Characterization of Three Types of $\delta$-Endotoxin Genes in Transformant, Bacillus thruingiensis PT0529 (형질전환체, Bacillus thuringiensis PT0529내에서 세가지 내독소 단백질 유전자들의 발현 특성)

  • 박현우;제연호
    • Journal of Sericultural and Entomological Science
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    • v.37 no.2
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    • pp.176-180
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    • 1995
  • To characterize expression and formation of three type crystal proteins in transformant, Bacillus thruingiensis PT0529 was analysed by transmission electron microscope and SDS-PAGE according to growth. The results showed that the introduced crystal protein genes, rcyIVD and cytA, were well expressed at earlier stage than resident crystal proteins were also expressed with their own morphology. However, resident crystal protein of B. thuringiensis PT0529 was smaller than that of wild type B. thuringiensis NT0423, suggesting that resident crystal protein production was interfered with introduced two type crystal protein genes.

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Development of Screening Method for the Soluble Recombinant Protein using β-Lactamase as a Fusion Partner (β-Lactamase 접합 단백질 발현 시스템을 이용한 가용성 재조합 단백질 탐색 기술 개발)

  • Lee, Jae-Hun;Hwang, Bum-Yeol;Kim, Byung-Gee;Lee, Sun-Gu
    • Korean Chemical Engineering Research
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    • v.47 no.5
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    • pp.624-629
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    • 2009
  • It is the most important step to screen soluble and insoluble proteins when we attempt to improve the solubility of recombinant proteins through directed evolution approach. Here we show that the solubility of a recombinant protein in vivo can be examined by expressing the recombinant protein with beta-lactamase as a fusion partner. First we constructed an expression system which can produc a fusion protein with the C-terminal of beta-lactamase. Two soluble proteins, i.e. adenine deaminase and aspartate aminotransferase, and insoluble GlcNAc-2-epimerase were cloned into the developed expression vector, respectively. We investigated the effect of the expression of the three recombinant fusion proteins on the growth of E. coli, and confirmed that the solubilities of the recombinant proteins correlated with cell growth rates.

Expression and Purification of Toll-like Receptor 9 Cytoplasmic Domain in Pichia patoris (Pichia pastoris로부터 Toll-like Receptor 9의 세포 내 도메인 단백질의 발현과 순수분리 정제)

  • Lee Kyun-Young;Lee Kon-Ho
    • Journal of Plant Biotechnology
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    • v.32 no.4
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    • pp.269-273
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    • 2005
  • Toll-like receptors (TLR) are important components of innate immunity in the defense against pathogens. TLRs recognize pathogen-associated common molecular patterns. TLRs are similar to the receptors involved in defense responses in plants. TLR protein is a type 1 membrane protein, consisting of an extracellular domain containing leucine-rich repeats and a cytoplasmic domain. The cytoplasmic domain delivers ligand recognition signals that result in production of anti-microbial agents. The cytoplasmic domain (amino acid 858-1032) of toll-like receptor 9 has been expressed using methylotrophic yeast Pichia pastoris. The protein expression was confirmed by Western-blot, N-terminal sequencing and MALDl-TOF mass spectrometry. The proteins have been purified by nickel affinity, cation exchange and gel-filtration chromatography.

Application of Data Cube to Identify Differentially Expressed Proteins by Disease (질병 의존 단백질 도출을 위한 데이터 큐브의 응용)

  • 김단비;이원석
    • Proceedings of the Korean Information Science Society Conference
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    • 2004.04b
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    • pp.268-270
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    • 2004
  • 주어진 셀이나 조직에 발현된 단백질 프로파일의 구조적인 분석을 다루는 단백질체학(Proteomics) 연구에 있어서, 질병에 대한 마커 단백질(marker proteins)을 도출(identification)하는 것은 핵심 논점 중 하나이다. 수십 개의 샘플로부터 추출한 셀이나 조직 내에는 수많은 단백질이 포함되어 있으며, 존재하는 단백질의 질병에 의한 발현량(expression level) 변화 및 임상 특성에 의한 영향을 분석하기 위해서 데이터베이스와 데이터 마이닝 기술의 활용이 효과적이다. 본 논문에서는 질병 일 임상 특성에 따른 단백질의 발현량 변화를 분석하기 위한 OLAP 데이터 큐브(Data cube)의 응용 방법과 단백질 데이터의 분석에 적합한 척도(measure)를 제안하고, 유효성을 보인다.

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Expression Pattern of RB and p53 Proteins and its Correlation with Prognosis in Primary Lung Cancer (원발성 폐암에서 종양억제유전자 RB와 p53 단백질 발현양상과 예후와의 상관관계)

  • 이상용;허혜경;최필조;우종수;홍숙희
    • Journal of Chest Surgery
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    • v.29 no.11
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    • pp.1223-1231
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    • 1996
  • Immunohistochemical stains for RB and p53 tumor suppressor gene products were performed on 72 cases of resected primary lung cancer tissues to study the correlation between their expressions and the histologic types, the clinical stage, and the survival rate. The results were as follows. 1. The RB protein was altered or absent in 38 cases (52.8%), and the mutant p53 protein was detected in 35 cases (48.6%). 2. The incidences of RB and p53 protein expression were significantly different among the histologic types (p<0.05) but were not correlated with the clinical stages of lung cancer (p>0.05). 3. The two year survival rate of patients with alteration of both RB and p53 genes (RB-/p53+) was 22. 4%, and that with no alteration of both genes (RB+/p53-) was 63.1%. This difference was statistically significant (p=0.01). 4. It was shown that alteration of RB protein greatly affects the prognosis of lung carcinoma by multivariate analysis of prognostic factors. The presence or absence of RB and mutant p53 protein in tumor cells is closely related to the survival of primary lung cancer patients, and it is suggested that RB gene expression is an independent prognostic factor of primary lung cancer.

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Prediction of epigenetic carcinogenesis based on protein network (단백질 네트워크 기반 후성유전학적 암 발생 기전 예측)

  • Jin, Hye Jeong;Lee, Jihoo;Kim, Hak Yong
    • Proceedings of the Korea Contents Association Conference
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    • 2016.05a
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    • pp.191-192
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    • 2016
  • DNA 염기서열 자체에는 변화가 없으나 크로마틴의 변형을 통하여 유전자의 발현 양상이 변하는 현상을 후성유전이라 한다. 최근에 이런 후성유전학적 변이가 암 발생과 밀접한 연관이 있는 것으로 알려졌다. 본 연구에서는 암 관련 단백질과 암 관련 후성유전 단백질 상호작용 네트워크를 통하여 암과 후성 유전적 관계를 분석하고자 하였다. 먼저 상호작용 네트워크를 기반으로 허브에 해당하는 히스톤 변형 단백질 20개를 추출하였다. 추출한 20개 단백질을 KEGG pathway에 적용하여 암 관련 단백질과의 상관관계를 분석하였다. 암 관련 단백질 발현양상을 확인할 수 있는 Expression Atlas로부터 발현이 증가하거나 감소하는 단백질을 분류하고, 발현 정보를 KEGG pathway 위에 있는 단백질에 적용함으로써 후성유전학적 암 발생 기전을 도출하였다.

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Expression of Antibacterial Protein, Nuecin, Using Baculorivus Expression Vector System in Bm5 Insect Cell and Bombyx mori (누에 배양세포(Bm5) 및 생체에서 베큘로바이러스 발현계를 이용한 누에신 단백질 발현 특성)

  • 윤은영;구태원;황재삼;김상현;강석우;김근영;진병래
    • Journal of Sericultural and Entomological Science
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    • v.44 no.2
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    • pp.69-73
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    • 2002
  • For the practical use of nuecin protein, we tried to overexpress nuecin using Bm5 insect cell and Bombyx mori. We inserted nuecin cDNA into pBm10po1-Xa vector derived from B. mori nuclear polyhedrosis virus (BmNPV), and expressed in Bm5 cells and B. mori respectively. SDS-PAGE and Northern blot analysis showed an expressed of the protein when baculovirus expression vector system (BEVS) was used. The amount of intracellular protein is abundant, but the amount of extracellular protein is poor. The results suggest that the biologically active nuecin protein produced by using BEVS is poor because incresed level of misfolded nuecin by the strong promoter, polyhedrin and p 10 of BEVS, decrease the level of free chaperons and foldases by binding with them.

Proteomic Changes in Odae Polished White Rice Grown at Different Cultivation Conditions (재배환경에 따라 변화하는 오대벼 백미의 단백질체 분석)

  • Lee, Ju-Young;Lee, Jin-Woo;Kim, Young-Ran;Yeom, Yu-Jin;Lim, Jin-Kyu
    • Journal of Applied Biological Chemistry
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    • v.55 no.2
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    • pp.79-83
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    • 2012
  • It has been known that the proteome profiles in the period of growth and development of rice are changed by the growth conditions including temperature, soil, and fertilization. In this study, the proteome profiles of Odae polished white rice grown in Chulwon and Chilgog were compared on 2-dimensional(D) gels. The differentially expressed proteins were selected from the 112 identified total proteins and classified into functional groups. The most significantly differentially expressed proteins were stress responsive proteins; Ent-kaur-16-ene synthase, which is responsible for synthesizing a plant hormone gibberellin, was expressed in Chulwon rice and heat shock proteins were in Chilgog rice, respectively. Xylanase inhibitor protein, which inhibits the enzyme xylanase produced by pathogenic fungi and Bacilli, was expressed significantly high in Chilgog rice grown at high temperature. Differential expressions of transporter proteins were observed both in Chulwon and Chilgog rice. Regarding the facts that Chilgog rice contained relatively higher amount of proteins than Chulwon rice and Chulwon rice showed large number of proteins were differentially expressed, it can be concluded that different cultivation conditions could change the protein expression profiles in rice in various ways, including elevation of protein amount or differential expressions of specific proteins, etc. The results suggest that the characteristics of the profiles of the proteome in the polished white rice are definitely changed by the environmental factors including high temperature. The results can be utilized for the development of the proper cultivation conditions for the production of high quality rice with good palatability.

Identification of differentially expressed proteins in the bacterial biofilm (세균성 바이오필름-특이 발현 단백질의 규명)

  • Kang, Chi-Dug;Choi, Jeam-Il
    • Journal of Periodontal and Implant Science
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    • v.35 no.2
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    • pp.271-275
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    • 2005
  • 본 연구는 치주낭에 biofilm형태로 부착되어 질환을 유발시키고 항생제 빚 항균제에 저항을 일으키는 세균 독성요소를 규명하기 위해 시행된 기초연구이다. 치주질환의 주 병원균의 하나인 Porphyromonas gingivalis 381 biofilm의 세포외막에 특이하게 발현되는 단백질을 규명하기 위한 기초적인 자료를 얻기 위해 시행하였다. Porphyromonas gingivalis 381을 통상적인 세균 배양용 broth를 사용하여 혐기성 세균 배양기로 24시간 배양한 것을 대조군으로 하고, tissue culture plate를 이용하여 혐기성 배양조건 하에서24시간동안 biofilm을 형성하여 실험군으로 설정하였다. 세균을 수획하여 세포외막을 분리하고 isotonic isoelectric focusing을 시행한 결과 주로 약 20-30 kilodaltons에 해당하는 수종의 세균세포막 단백질이biofilm으로 배양한 세균에서 더 상승적으로 발현됨이 관찰되었고, 상이한 수종의 단백질도 planktonic culture broth로 배양한 세균에서 다 상승적으로 발현됨을 관찰할 수 있었다. 이것은 세균의 배양조건과 환경에 따라 그 외막 단백질이 서로 다르게 발현됨을 입증하는 기초적인 자료로서 향후 단백질의 동정과 성격을 규명하는 근간 실험으로 추진할 계획이다.

In Sitilico Protein Sequencing Based on Mass Spectrometry Using Multiple Pretenses (다중 효소를 이용한 질량분석기법에 기반한 단백질의 아미노산 서열 분석)

  • 문석현;이도헌;이광형
    • Proceedings of the Korean Institute of Intelligent Systems Conference
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    • 2002.12a
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    • pp.473-477
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    • 2002
  • 세포내에서 특정 단백질이 합성되어 이용되는 것을 단백질의 발현이라 한다. 이러한 단백질의발현을 조사하는 작업은 세포내 대사과정을 밝혀내는 데 있어서 매우 중요한 역할을 담당하고 있다. 단백질의 발현을 조사하기 위해서는 세포로부터 추출하여 정제한 단백질이 어떤 단백질인지를 확인하는 작업이 필요한데 현재로써는 확인하고자 하는 단백질 효소로 분해하여 분해된 조각들의 질량을 측정하여 기존에 알려진 단백질들을 분해했을 때 이론상 나을 수 있는 조각들의 무게와 비교하여 가장 근접한 단백질을 찾아내는 질량분석기법(mass Spectrometry)이 널리 사용된다. 그러나 이 방법은 확인하고자 하는 단백질의 아미노산 서열이 알려져 있을 경우에만 사용할 수 있다는 한계점을 가지고 있다. 본 논문에서는 이러한 한계를 계산적인 방법으로 극복하고자 동일단백질을 여러가지 효소로 분해하여 나오는 조각들의 질량을 측정하고 이들을 조합하여 원래 단백질의 아미노산 서열을 알아낼 수 있는 알고리즘을 제안한다.