• 제목/요약/키워드: 2-Dimensional gel electrophoresis

검색결과 216건 처리시간 0.027초

A Proteomic Approach to Study msDNA Function in Escherichia coli

  • Jeong, Mi-Ae;Lim, Dongbin
    • Journal of Microbiology
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    • 제42권3호
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    • pp.200-204
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    • 2004
  • Retron is a prokaryotic genetic element that produces multicopy single-stranded DNA covalently linked to RNA (msDNA) by a reverse transcriptase. It was found that cells producing a large amount of msDNA, rather than those that did not, showed a higher rate of mutation. In order to understand the molecular mechanism connecting msDNA production to the high mutation rate the protein patterns were compared by two dimensional gel electrophoresis. Ten proteins were found to be differentially expressed at levels more than three fold greater in cells with than without msDNA, nine of which were identified by MALDI TOF MS. Eight of the nine identified proteins were repressed in msDNA-producing cells and, surprisingly, most were proteins functioning in the dissimilation of various carbon sources. One protein was induced four fold greater in the msDNA producing cells and was identified as a 30S ribosomal protein S2 involved in the regulation of translation. The molecular mechanism underlying the elevated mutation in msDNA-producing cell still remains elusive.

Proteomic Analysis of Drought Stress-Responsive Proteins in Rice Endosperm Affecting Grain Quality

  • Mushtaq, Roohi;Katiyar, Sanjay;Bennett, John
    • Journal of Crop Science and Biotechnology
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    • 제11권4호
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    • pp.227-232
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    • 2008
  • Drought stress is one of the major abiotic stresses in agriculture worldwide. We report here a proteomic approach to investigate the impact of post-fertilization drought on grain quality in rice seed endosperm (Oryza sativa cv. IR-64). Plants were stressed for 4 days at 3 days before heading. Total proteins of endosperm were extracted and separated by two-dimensional gel electrophoresis. Not many protein spots showed differential accumulation in drought-stressed samples. More than 400 protein spots were reproducibly detected, including three that were up-regulated and five down-regulated. Mass spectrometry analysis and database searching helped us to identify six spots representing different proteins. Functionally, the identified proteins were related to protein synthesis and carbohydrate metabolism, such as Granule-Bound Starch Synthase (GBSS, Wx protein), which is thought to play a very important role in starch biosynthesis and quality, a very crucial factor in determining rice grain quality.

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Microbial Diversity Information Facility: Bacteriology Insight Orienting System (BIOS)

  • Shimura, Junko;Shimiz, Hideyukiu;Tsuruwaka, Keiji;Moritani, Yukimitsu;Miyazaki, Kenji;Tsugita, Akira;Watanabe, Makoto M.
    • 한국미생물생명공학회:학술대회논문집
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    • 한국미생물생명공학회 2000년도 Proceedings of 2000 KSAM International Symposium and Spring Meeting
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    • pp.135-141
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    • 2000
  • Global Biodiversity is common interest of humans for better health and sustainable development of the society. To provide access and analysis on microbial diversity information, Bacteriology Insight Orienting System (BIOS) has been developed. BIOS contains 6402 species and subspecies names of bacteria and archaea, 2606 names of cyanobacteria by March 2000. BIOS of which web based analytical tool provides windows to compare the results of phylogenetic analysis based on 16S rDNA sequence and the results of cluster analysis on proteome profiling. The sequence data and 2 dimensional gel electrophoresis analysis data were accumulated in BIOS database content for cyanobacteria reclassification and taxonomy. (BIOS URL: http.://www-sp2000ao.nies.go.jp/bios/index.html).

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전리수를 사용한 이차원 전기영동법 (2-Dimensional gel electrophoresis by using electrolyzed water)

  • 민병술;이윤배;이종권;류근걸;이미영
    • 한국산학기술학회:학술대회논문집
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    • 한국산학기술학회 2004년도 추계학술대회
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    • pp.259-261
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    • 2004
  • 일반 증류수로 제조된 완충용액으로 미세조류를 파쇄하였을 때 미세조류의 두꺼운 세포벽에 의해 단백질 추출이 효율적으로 이루어지지 않는다. 따라서 일차원 및 이차원 전기영동법을 사용하여 단백질 발현 양상을 분석할 때 효율이 떨어진다. 본 연구에서는 물의 클러스트가 작아 세포벽에 잘 흡수될 수 있다고 알려진 환원전리수를 사용하여 효율적으로 단백질을 추출하였고, 이차원 전기영동법으로 단백질 발현 양상을 확인하였다.

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질량분석기를 단백질 분석에 적용하기 위한 고성능액체크로마토그래피 최적조건 연구 (A study on the optimal HPLC condition for peptides complex analysis using mass spectrometry)

  • 권성원;박철홍
    • 분석과학
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    • 제16권1호
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    • pp.78-81
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    • 2003
  • Peptides separation in high performance liquid chromatography (HPLC) is very important for the analysis of total proteins using mass spectrometry rather than two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). In this study, we investigated the optimal HPLC condition of peptides for the use of mass fingerprinting. As a result of pursuing a combination of solvent additives for HPLC, water and acetonitile containing both 0.1% trifluoroacetic acid and 0.1% acetic acid respectively showed the most efficient resolution and sensitivity.

돼지에서 발정 주기 동안 자궁액 내의 단백질 패턴의 변화 (Change of Protein Patterns in Uterine Fluid during Estrus Cycle in Pigs)

  • 이연주;송은지;이상희;정희태;양부근;박춘근
    • Reproductive and Developmental Biology
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    • 제37권3호
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    • pp.103-108
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    • 2013
  • An uterus is female reproductive tract organ that affected estrus cycle. During a various changes occur at uterus in estrus cycle, one of them is body fluids secretion be called uterine fluid. Therefore, the objective of this study was to investigate the changes of protein patterns using two-dimensional gel electrophoresis in uterus fluids during the follicular and luteal phases in estrus cycle of pigs. In changes of protein spots were confirmed during the follicular and luteal phases. The 136 spots were expressed in follicular phase, the 57 spots of them showed reproducibility. On the other hand, the 140 spots were expressed in luteal phase, the 73 spots of them showed reproducibility. Also, spots expressed in follicular phase were number 69 and 94 spots and spots expressed in luteal phase only were number 156, 157, 184~187, 190 and 191 spots. The spots which of higher expression levels in the luteal phase than in follicular phase were number 76 and 79 spots. In conclusion, the spots expressed in follicular and luteal phases were confirmed with difference levels and these differences are function of RNA resolving, protein synthesis and cytoskeletal architecture.

非姙娠 및 姙娠한 女子의 血淸蛋白質 패턴의 比較 (The Comparison of Protein Patterns of Sera in Non-Pregnant and Pregnant Women)

  • Ha, Man-Joon;Park, Won-Chul
    • 한국동물학회지
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    • 제29권2호
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    • pp.86-106
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    • 1986
  • 사람의 혈액중 혈청으로써 임신의 진행에 따른 단백질의 변화 양상을 조사함에 있어서 정상인 남자, 비임신 여자, 임신한 여자 및 분만 직후 여자의 혈청을 SDS/polyacrylamide gel 전기영동과 2차원 전기영동 및 아미노산분석을 시도하였다. 각 실험 방법에 의해 분석된 결과는 다음과 같다. 1. SDS/polyacrylamide gel 전기영동에 의해 분자량 약 10,000에서 110,000 dalton 사이의 정상인 남자와 비임신 여자를 비교하였을 때의 숫적 양상은 서로 동일하였으나 bands 3 (22,000 dalton)과 6(39,000)등은 남자에 있어서 여자보다 적은 양으로 나타났다. 비임신 여자와 2주 간격으로 혈액을 채취한 임신한 여자의 혈청단백질 패턴을 비교하면 여러 band들의 양적인 증감이 관찰되었다. 임심한 여자의 단백질은 비임신에 비해 band 3(22,000)이 16주까지는 비임신의 경우와 거의 비슷한 양상을 보였으나 18주부터 임신 말기까지는 단백질의 양이 계속하여 감소되었다. 반면 bands 4(24,000), 9(69,000), 10(70,000), 12(80,000), 13(83,000), 14(86,000), 15(91,000) 및 16(94,000)은 임신이 진행됨에 따라 일반적으로 증가되나 그 이후에는 약간씩 감소하였다. 분만 직후의 혈청단백질의 패턴을 임신 말기와 비교하였을 때 bands 12(80,000), 15(91,000) 및 16(94,000)들의 양이 비교적 증가하였고 여아를 분만한 경우 남아를 분만한 경우보다 bands 4(24,000), 7(51,000) 및 10(70,000)들의 단백질 양이 많음이 관찰되었다. 2. 2차원 전기영동으로 남자와 비임신 여자의 혈청단백질 패턴을 비교하면 spot a(22,000)의 3개의 spot가 남자에 있어서는 나타나지 않았고, spotc(39,000)의 군은 남자에 있어 여자보다 농도가 매우 낮았다. 임신한 여자에 있어 albumin이 임신 10주와 12주에 매우 감소하였고 그 이후에 다시 회복되었다. 그리고 spot f(70,000)는 임신 10주에 매우 감소 되었다가 그 이후에 증가되었다. 3. 각 군들의 아미노산을 분석한 결과 임신한 여자에 있어서 전반적으로 glutamic acid, lysine, aspartic acid, leucine 및 valine이 대체로 많은 양으로 나타났고, methionine, isoleucine 및 glycine이 대체로 적은 양을 가지는 것으로 나타났다. 전체 아미노산의 양은 임신중기에 현저히 증가하였다가 말기에는 중기에 비해 감소되었다. 이상의 결과를 볼 때 임신기간 중에는 모체의 혈청을 구성하는 단백질이 특정한기에 따라 증가하므로 임신한 여자의 혈청단백질을 조사함으로써 임신의 각기에 나타나는 독특한 단백질 양상 변화를 밝혀내는 동시에, 앞으로 좀더 연구함으로써 태아의 성을 단백질 패턴으로 명확히 구별해 낼 수 있을 것으로 생각된다.

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Identification of Granule Bound Starch Synthase (GBSS) Isoforms in Wheat

  • Seo, Yong-Weon;Hong, Byung-Hee;Ha, Yong-Woong
    • 한국작물학회지
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    • 제43권2호
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    • pp.89-94
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    • 1998
  • Granule bound starch synthase (GBSS), also known as the '"waxy protein'", is responsible for the synthesis of amylose in the amyloplasts of cereal crops. In hexaploid wheat (Triticum aestivum L.), GBSS is involved in amylose synthesis and rolls as an important factor to determine flour quality and end-use quality in food products. Genes on three Wx loci have been found to encode GBSS in common wheats. We developed techniques for the purification and separation of GBSS in wheat. Three major GBSS isoforms, which were encoded by the genes on three loci, Wx-A1, Wx-B1, and Wx-D1 migrating differently by one dimensional SDS-po-lyacrylamide gel electrophoresis (1D SDS-PAGE), were identified. GBSS from 66 Korean hard and soft winter wheats were purified and determined for their Wx loci and four of them were identified possessing a null allele either at the Wx-A1 and Wx-B1 loci. With help of identification of three GBSS isoforms using 1D SDS-PAGE system, we are able to identify and monitor Wx gene expressions in breeding materials for developing waxy or partial waxy wheats without experiencing consecutive selecting generations.cting generations.

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Two-Dimensional Reference Map of Schizosaccharomyces pombe Proteins (Update)

  • Kim, Sun-Kyung;Won, Mi-Sun;Sun, Nam-Kyu;Jang, Jae-Won;Lee, Seung-Hee;Shin, Hee-Young;Song, Kyung-Bin
    • Journal of Microbiology and Biotechnology
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    • 제16권10호
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    • pp.1499-1512
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    • 2006
  • Based on the first 2D reference map of the fission yeast Schizosaccharomyces pombe protein reported previously, we expanded and updated the map using narrower pI ranges. In this paper, 240 protein spots were identified on our reference map. In the pI 4-7 range, 144 spots corresponding to 86 different proteins were identified. In the pI 6-9 range, 43 spots corresponding to 35 different proteins were identified. Fifty-three new spots corresponding to 39 different proteins were further identified in the pI 5-6 range.

Identification of Bovine Pregnancy-Specific Whey Proteins using Two-Dimensional Gel Electrophoresis

  • Han, Rong-Xun;Choi, Su-Min;Kim, Myung-Youn;Quan, Yan Shi;Kim, Baek-Chul;Diao, Yun Fei;Koqani, Reza;Park, Chang-Sik;Jin, Dong-Il
    • Reproductive and Developmental Biology
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    • 제32권4호
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    • pp.255-261
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    • 2008
  • The early diagnosis of bovine pregnancy is an essential component of successful reproductive planning on farms, because lack of bovine pregnancy over the long term results in reproductive failure and low milk yield-the latter of which is a special concern on dairy farms. This study was designed to identify early pregnancy-specific whey proteins in bovine, by comparing milk samples collected from cattle during pregnancy (Days 30 and 50) and from non-pregnant cattle. In this study, differentially expressed proteins in five pregnant and five non-pregnant Holstein dairy cattle were investigated and compared, using proteomics analysis. The first dimension was applied to a pH $3.0{\sim}10.0$ strip, by loading a 2-mg milk protein sample. After the second-dimension separation was performed, the gels were stained with colloidal Coomassie brilliant blue. The stained gels were scanned and the images were analyzed, to detect variations in protein spots between non-pregnant and pregnant cattle milk protein spots, using ImageMaster, this was followed by analysis with MALDI TOF-MS. Analysis of the 2-DE gel image resulted in a total of approximately $500{\sim}600$ protein spots, of which 12 spots were differentially expressed, six spots were up-regulated, and four spots were down-regulated; two spots were identified as pregnancy-specific proteins. These proteins were identified as lactoferrin, NA-DH dehydrogenase subunit 2, albumin, serum albumin precursor and transferrin. Our results via 2-D PAGE analysis revealed composite profiles of several milk proteins related to early bovine pregnancy, implying the possible use of these milk proteins in the early detection of bovine pregnancy.