• Title/Summary/Keyword: size exclusion

Search Result 245, Processing Time 0.026 seconds

High Yield Bacterial Expression and Purification of Active Cytochrome P450 p-coumarate-3-hydroxylase (C3H), the Arabidopsis Membrane Protein (대장균 시스템을 이용한 Arabidopsis 막 단백질 cytochrome P450 p-coumarate-3hydroxylase (C3H) 활성형의 과발현 및 분리정제)

  • Yang, Hee-Jung;Kim, Wan-Yeon;Yun, Young-Ju;Yoon, Ji-Won;Kwon, Tae-Woo;Youn, Hye-Sook;Youn, Bu-Hyun
    • Journal of Life Science
    • /
    • v.19 no.8
    • /
    • pp.1039-1046
    • /
    • 2009
  • The cytochrome P450s (P450s) metabolizing natural products are among the most versatile biological catalysts known in plants, but knowledge of the structural basis for their broad substrate specificity has been limited. The activity of p-coumarate 3-hydroxylase (C3H) is thought to be essential for the biosynthesis of lignin and many other phenylpropanoid pathway products in plants however, all attempts to express and purify the protein corresponding C3H gene have failed. As a result, no conditions suitable for the unambiguous assay of the enzyme are known. The detailed understanding of the mechanism and substrate-specificity of C3Hdemands a method for the production of active protein on the milligram scale. We have developed a bacterial expression and purification system for the plant C3H, which allows for the quick expression and purification of active wild-type C3H via introduction of combinational mutagenesis. The modified cytochrome P450 C3H ($C3H_{mod}$) could be purified in the absence of detergent using immobilized metal affinity chromatography and size exclusion chromatography following extraction from isolated membranes in a high salt buffer and catalytically activated. This method makes the use of isotopic labeling of C3H for NMRstudies and X-ray crystallography practical, and is also applicable to other plant cytochrome P450 proteins.

Glomus Tumor in Soft Tissue (연부 조직에 발생한 사구종)

  • Kim, Do-Yeon;Lee, Soo-Hyun;Kim, Min-Ju;Shin, Kyoo-Ho
    • The Journal of the Korean bone and joint tumor society
    • /
    • v.15 no.1
    • /
    • pp.34-43
    • /
    • 2009
  • Purpose: Glomus tumors are rare benign vascular tumors, usually located in the skin or soft tissue of extremities. Approximately 30-50% of glomus tumors occur in subungal area, but glomus tumors have been described in every location even where glomus bodies are not or rarely present. The purpose of this study was to identify clinical, histologic and MRI characteristics of soft tissue glomus tumor. Materials and Methods: Between 1993 and 2008, eight patients underwent surgery of soft tissue Glomus tumor at our institution. Exclusion criteria were patients with Glomus tumors in digits, stomach, trachea and glomus tympanicum. We analyzed medical records, interviews, physical examinations, MR findings and histolocial types retrospectively. Results: There were four men and four women. The mean age was fourty-seven years. The mean prevalence time was eight-point-nine years. In the classic triad of symptoms, all eight patients had pain and tenderness. Two patients complained of cold sensitivity. Two showed skin color changes. After surgery, two showed symptom improvement (VAS $9^{\circ}{\rightarrow}8$, $8^{\circ}{\rightarrow}5$) and?six showed complete disappearance of symptoms. Slightly symptom improvemented (VAS $9^{\circ}{\rightarrow}8$) one had additional surgery two times after first surgery due to relapse/remaining Glomus tumor. The mean size was 13.9 mm. In histology, six were 'solid glomus tumor', one was a mixture of 'solid glomus tumor' and 'lomangioma' and one was 'malignant glomus tumor'. MR findings showed isointense signal on T1 image, high signal on T2 image and strong enhancement on the Gadolinium enhanced image. Conclusion: Glomus tumor has low recurrence rate and malignant change, rapid diagnosis and surgical excision is critical in treatment to prevent unnecessary pain of patient.

  • PDF

Purification and Characterization of Sulfated Polysaccharide Isolated from Hot Water Extract of Pachymeniopsis elliptica (Pachymeniopsis elliptica의 열수 추출물로부터 분리한 함황 다당류의 정제 및 특성)

  • Lee, Sun-Hee;Jun, Woo-Jin;Yu, Kwang-Won;Chun, Hyug;Shin, Dong-Hoon;Hong, Bum-Shik;Cho, Hong-Yon;Yang, Han-Chul
    • Korean Journal of Food Science and Technology
    • /
    • v.32 no.5
    • /
    • pp.1191-1197
    • /
    • 2000
  • In the preliminary study, we investigated the anti-complementary activities of 62 extracts from Korean edible seaweeds. Of those, Pachymeniopsis elliptica showed the highest anti-complementary activity. Therefore, it was purified as follows; i) PE-1 by ethanol precipitation, ii) PE-1-C by ultrafiltration, iii) PE-1-CIV by DEAE-Toyopearl 650C, and iv) PE-1-CIV-ii by Sepharose CL-6B. The purified compound, PE-1-CIV-ii, was the complexed homogeneous polysaccharide (molecular mass: 780 kDa) with 82.9% of anti-complementary activity. Also, it contained a significant amount of sulfate group (30.5%), which indicated it as a sulfated algal polysaccharide. Its structural monosaccharides were galactose (44.3%), 3,6-anhydrogalactose (34.0%), glucose (8.2%), fucose (5.4%), xylose (5.2%) and rhamnose (2.9%). After the treatment of periodate on a sample, a significant decrease in anti-complementary activity was found, which was a characteristic of bioactive polysaccharides. And-tumor activity of PE-1-A, B and C was tested in the sarcoma-180 solid tumor model. The PE-1-C with the largest molecular mass (more than 300 kDa) showed 81% of inhibition on the solid tumors, suggesting that the anti-complementary activity was, at least in part, related to anti-tumor activity. Based upon these results, the purified polysacchardes could be an immunopotentiator in vivo.

  • PDF

A Comparison of Discriminating Powers between 13 Microsatellite Markers and 37 Single Nucleotide Polymorphism Markers for the Use of Pork Traceability and Parentage Test of Pigs (돼지 개체식별 및 친자감별을 위한 13 microsatellite marker와 37 single nucleotide polymorphism marker 간의 효율성 비교)

  • Lee, Jae-Bong;Yoo, Chae-Kyoung;Jung, Eun-Ji;Lee, Jung-Gyu;Lim, Hyun-Tae
    • Journal of agriculture & life science
    • /
    • v.46 no.5
    • /
    • pp.73-82
    • /
    • 2012
  • Allele information from the analysis of the 13 microsatellite (MS) markers, were classified into the $F_0$, $F_1$ and $F_2$ generations, and probabilities of the same individual emergency in each generation was calculated. As a result, the 13 MS markers showed an estimate of $3.84{\times}10^{-23}$ on the premise of the randomly mated group of $F_2$, which implies that the same individuals may emerge by the use of 37 kinds of SNP markers. In this study, the experimental pigs were intercross between only 2 breeds (Korean native pig and Landrace). In addition, the success rate of paternity tests was analyzed on the whole group, by the use of the 13 MS markers and 37 SNP markers. As regards the exclusionary power of the second parent ($PE_{pu}$), MS markers and SNP markers showed 0.97897 and 0.99149, respectively. In relation to the parent exclusion power of both parent (PE), MS markers and SNP markers showed 0.99916 and 0.99949, respectively. In the case of the estimate to identify parental candidates that had the highest probability ($PNE_{pp}$), the two showed 1.00000 all. The Korean pig industry tends to mass produce hogs with limited numbers of alleles in limited parents. Such being the case, there is a need to organize a marker, for which it is imperative to find markers with high efficiency and high economic feasibility of the characteristics of DNA markers, sample size, the accuracy and expenses of genotyping cost, the manageability of data and the compatibility among analysis systems.

Experimental Studies on the Antitumor Effects of Jinryungtang Gagambang Extract (진령탕가감방의 항종양효과(抗腫瘍效果)에 관(關)한 실험적(實驗的) 연구(硏究))

  • Jeong, Jun-Tak;Moon, Goo;Moon, Suk-Jae
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
    • /
    • v.4 no.1
    • /
    • pp.37-53
    • /
    • 1998
  • The sprig of Jinryungtang Gagambang has been used for curing as a traditional medicine without any experimental evidence to support the rational basis for their clinical use. This experiment was carried out to evaluate the possible therapeutic or antitumoral effects of Jinryungtang Gagambang extract against cancer, and to study some mechanisms responsible for its effect. The cytotoxic and antitumor effects were evaluated on human cell liens (A549, hep3B, Caki-1, Sarcoma 180) after exposure to Jinryungtang Gagambang extract using in ILS, colony forming efficency and SRB assay which were regarded as a valuable method for cytotoxic and antitumor effects of unknown compound on tumor cell lines. The results obtained in this studies were as follows. 1. As a result of exposure to Jinryungtang Gagambang extract, the proliferation of A549, hep3B, Caki-1, good correlations were shown from the results of SRB assay and those of clogenetic assay. 2. The oral administration of Jinryungtang Gagambang extract showed significant effects of increase of MST(mean survival time) and ILS(increased life span) depending on the increasing concentration. 3. Against squamous cell carcinoma induced by MCA, Jinryungtang Gagambang decreased not only the frequency of tumor production but also the number and weight of tumors per tumor bearing mice(TBM). Jinryungtang Gagambang also significantly suppressed the development of 3LL cell-implanted tumors by frequency and their size, and some developed tumors were regressed by the continuous treatment of Jinryungtang Gagambang extract into TBM. 4. Jinryungtang Gagambang extract also increased NK cell activities. According to the above results, it could be suggested that Jinryungtang Gagambang extract has prominent antiutmor effect.

  • PDF