• Journal of Plant Biology
• /
• 제38권2호
• /
• pp.143-151
• /
• 1995

Addition of plant growth hormones [0.01 mg/L NAA and 0.2mg/L benzyladenine (BA)] to a woody plant medium stimulated the adventitious bud formation of poplar explants during culture. Endogenous IAA content increased rapidly at the initial culture stage and then decreased, being followed by rapid increment again at the late culture. But the content of trans-zeatin riboside (t-ZR) increased continuously during the culture. Cytoplasmic soluble proteins were analyzed by one- and two-dimensional SDS-PAGE. Increased amount of 40 kD band was detected by one-dimensional electrophoresis using Coomassie Blue staining during the culture and two distinctive proteins whose mol wt is 40,000 were detected by two-dimensional electrophoresis using autoradiography and these proteins were synthesized continuously prior to the adventitious bud formation. When the midvein segments were transferred to the actinomycin D-containing medium, the spots of adventitious bud-related proteins(ABRPs) did not disappeared but weakened in intensity. So, it is concluded that genes coding for the ABRPs are regulated to some degree at the transcriptional level. Also, they were not observed in BA-free medium, suggesting that these proteins be regulated by cytokinin, which made then possible to form the adventitious bud.

• 한국컴퓨터정보학회논문지
• /
• 제14권7호
• /
• pp.41-47
• /
• 2009

단백질 연구에 획기적인 기여를 한 2차원 전기영동법에 의한 결과영상을 처리하기 위하여 일반적으로 제안되었던 기존의 영상처리기법들이 가지는 문제점 중 하나인 단백질을 나타내는 여러 개의 스팟들이 겹치고 포화되고 있는 복잡영역에서 단백질 스팟을 하나로 인식하거나 검출이 불가능하여 오류를 범하게 되는 문제점에 대한 해법을 제안한다. 복잡영역내의 각 각점의 누적 기울기를 산출하고 누적 기울기 영상을 워터쉐드 기법으로 영역 분할하여 스팟을 분리한다. 그 결과 스팟 분리에 있어서 기존의 기법들을 적용하였을 때 보다 더 우수하고 효율적인 결과를 나타내며, 2차원 전기영동 결과영상에 숨겨져 있었던 단백질 스팟을 더 많이 검출할 수 있고, 예측의 범위 또한 확장시킨다.

• International Journal of Industrial Entomology and Biomaterials
• /
• 제17권2호
• /
• pp.217-221
• /
• 2008

Dung beetle larvae at the $3^{rd}$ instar were injected with lipopolysaccaride and inducible proteins were examined within a pI level of 3-10 and a size level by proteomics, including 1-D SDS PAGE analysis and antibacterial assay. The immune infected larvae extracts provided seven protein bands in one-dimensional electrophoresis and its antibacterial activity also checked. Hemolymph protein from immune infected larvae of the dung beetle were separated by twodimensional gel electrophoresis and compared with those from native larvae. In 2-D gel electrophoresis, we detected 63 immune infected unique and 32 up-regulated proteins, and 36 proteins that were down-regulated or not present in treated gel. Ten protein spots from unique proteins and those presented as different level of abundance in infected and native larvae were specially expressed. These differentially expressed proteins were proposed to be involved in the defense mechanism against microorganism.

• 한국생물공학회:학술대회논문집
• /
• 한국생물공학회 2005년도 생물공학의 동향(XVII)
• /
• pp.781-781
• /
• 2005

• 한국생물공학회:학술대회논문집
• /
• 한국생물공학회 2005년도 생물공학의 동향(XVII)
• /
• pp.832-832
• /
• 2005

• BMB Reports
• /
• 제39권2호
• /
• pp.216-222
• /
• 2006

In the present study, we compared six different solubilization buffers and optimized two-dimensional electrophoresis (2-DE) conditions for human lymph node proteins. In addition, we developed a simple protocol for 2-D gel storage. Efficient solubilization was obtained with lysis buffers containing (a) 8M urea, 4% CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate), 40 mM Tris base, 65 mM DTT(dithiothreitol) and 0.2% carrier ampholytes; (b) 5M urea, 2M thiourea, 2% CHAPS, 2% SB 3-10 (N-decyl-N, N-dimethyl-3-ammonio-1-propanesulfonate), 40mM Tris base, 65 mM DTT and 0.2% carrier ampholytes or (c) 7M urea, 2M thiourea, 4% CHAPS, 65 mM DTT and 0.2% carrier ampholytes. The optimal protocol for isoelectric focusing (IEF) was accumulated voltage of 16,500 Vh and 0.6% DTT in the rehydration solution. In the experiments conducted for the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), best results were obtained with a doubled concentration (50 mM Tris, 384 mM glycine, 0.2% SDS) of the SDS electrophoresis buffer in the cathodic reservoir as compared to the concentration in the anodic reservoir (25 mM Tris, 192 mM glycine, 0.1% SDS). Among the five protocols tested for gel storing, success was attained when the gels were stored in plastic bags with 50% glycerol. This is the first report describing the successful solubilization and 2D-electrophoresis of proteins from human lymph node tissue and a 2-D gel storage protocol for easy gel handling before mass spectrometry (MS) analysis.

• 한국해양바이오학회지
• /
• 제2권1호
• /
• pp.68-72
• /
• 2007

넙치의 혈청 단백질체를 연구하기 위해 본 연구에서는 이차원전기영동 분석 시의 기본조건을 확립하였다. 넙치의 혈청단백질을 이차원전기영동 법으로 분석하기 위해서는 TCA 침전을 이용하여 혈청 내에 고농도로 존재하는 이온물질을 제거하는 전처리과 정이 반드시 요구되었다. 또한 분석 단백질의 농도는 $30{\mu}g$이 적당하였다. 넙치 10개체의 혈청단백질을 이용하여 총 51회의 이차원전기영동을 수행한 결과 1,8207개의 단백질로 이루어진 넙치 혈청단백질 표준 지도를 작성할 수 있었다.

• Mass Spectrometry Letters
• /
• 제3권1호
• /
• pp.10-14
• /
• 2012

Artificially oxidized cysteine residues in peroxiredoxin 6 (Prx6) were detected by electrospray interface capillary liquid chromatography-linear ion trap mass spectrometry after the preparation of two-dimensional gel electrophoresis (2D-GE). We used Prx6 as a model protein because it possesses only two cysteine residues at the 47th and 91st positions. The spot of Prx6 on 2D-GE undergoes a basic (isoelectric point, pI 6.6) to acidic (pI 6.2) shift by exposure to peroxide due to selective overoxidation of the active-site cysteine Cys-47 but not of Cys-91. However, we detected a tryptic peptide containing cysteine sulfonic acid at the 47th position from the basic spot and a peptide containing both oxidized Cys-47 and oxidized Cys-91 from the acidic spot of Prx6 after the separation by 2D-GE. We prepared two types of oxidized Prx6s: carrying oxidized Cys-47 (single oxidized Prx6), and other carrying both oxidized Cys-47 and Cys-91 (double oxidized Prx6). Using these oxidized Prx6s, the single oxidized Prx6 and double oxidized Prx6 migrated to pIs at 6.2 and 5.9, respectively. These results suggest that oxidized Cys-47 from the basic spot and oxidized Cys-91 from the acidic spot are generated by artificial oxidation during sample handling processes after isoelectric focusing of 2D-GE. Therefore, it is important to make sure of the origin of cysteine oxidation, if it is physiological or artificial, when an oxidized cysteine residue(s) is identified.

• 정보처리학회논문지B
• /
• 제17B권1호
• /
• pp.79-84
• /
• 2010

두 개의 이차원 단백질 전기영동 영상에서 한쪽 영상의 각 반점이 다른 영상의 어느 반점과 일치하는 지 알아내는 일은 단백질의 생성과 소멸, 변이 등을 알 수 있게 하는 중요한 일이다. 단백질 반점은 주로 2차원 전기영동에 의해 분리된다. 이 과정은 조직의 상태, 실험 조건 등에 따라 같은 단백질이라도 그 위치가 조금씩 다르게 된다. 각 반점을 정합하여 보면 균일하지 않은 비선형 변환 관계를 이룬다. 그러나 국지적으로 보면 반점 사이의 토폴로지가 유지되는 것을 볼 수 있다. 본 연구는 이러한 토폴로지의 유지에 착안하여 반점끼리 정합하는 방법을 제안한다. 토폴로지의 유사성을 비교하기 위하여 이웃한 반점과의 거리와 각도를 비교하였다. 실험을 통하여 제안한 방법이 우수함을 보였다.

• 미생물학회지
• /
• 제54권3호
• /
• pp.222-227
• /
• 2018

CD1 분자는 결핵균 유래 지질항원 발현하는 단백질이며, 특히 수지상세포(dendritic cells)가 결핵균 감염시에 발현이 점차 감소함을 관찰하였다. 이는 결핵균의 사균이나 항원만으로는 관찰되지 않는 결과였다. 2차원 전기영동(2D electrophoresis)을 통하여 CD1b 의 인산화를 관찰하였고 이러한 현상이 식세포작용과 연관됨을 확인하였다.