生物物理化学
Online ISSN : 1349-9785
Print ISSN : 0031-9082
ISSN-L : 0031-9082
42 巻, 3 号
選択された号の論文の9件中1~9を表示しています
  • 浅川 順一
    1998 年 42 巻 3 号 p. 145-154
    発行日: 1998/08/15
    公開日: 2009/03/31
    ジャーナル フリー
    A vertical giant gel two-dimensional electrophoresis (2-DE) system has been developed and applied to the restriction landmark genomic scanning (RLGS; 2 DE of end labeled genomic DNA fragments). A unique gel holder for the first-dimension agarose disc gel was made from teflon-tubing and glass tube. The development of the disc gel system resulted in high-resolution separation of DNA fragments with enhanced reproducibility. A new device for the in-gel restriction enzyme digestion of the fractionated DNA fragments was also made from teflon-tubing. After optimization of the sample preparation and electrophoresis conditions, the gel quality reached a level such that the electrophoresis patterns derived from a single DNA sample gave distribution patterns of spots able to be superimposed. A protocol and the equipment for the 2-DE are described.
  • 島崎 洋次, 真鍋 敬
    1998 年 42 巻 3 号 p. 155-159
    発行日: 1998/08/15
    公開日: 2009/03/31
    ジャーナル フリー
    Using microscale multisample two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) in the absence of denaturing agents, eight protein samples can be simultaneously analyzed and compared. In the present study, this method was applied for the analysis of chicken serum, chicken egg yolk and chicken egg albumen proteins, and the 2-D PAGE pattern of chicken yolk and albumen were compared with that of chicken serum. The spot locations (pI and Mr) of yolk proteins resembled with that of chicken serum, but those of albumen proteins were considerably different from those of chicken serum. Employing the techniques of electrophoretic blotting and immunochemical staining, immunoglobulin G (IgG), third component of complement (C3) and transferrin were identified in the patterns of chicken serum and yolk proteins.
  • 星野 忠, 松下 誠, 入野 勤, 熊坂 一成, 河野 均也, 菰田 二一
    1998 年 42 巻 3 号 p. 161-167
    発行日: 1998/08/15
    公開日: 2009/03/31
    ジャーナル フリー
    小腸ALPには少なくとも通常の小腸ALP, 小腸様ALP, 血液型依存性高分子小腸ALP, 胎児小腸ALPの4アイソフォームが知られている. PAG電気泳動法ではこれら4アイソフォームの小腸ALPを明確に判別することは可能であるが, セルロースアセテート膜およびアガロースゲル電気泳動法では各種小腸ALPの泳動位置に関する詳細な報告がないので今回の検討を実施した. ALPアイソザイムパターン, Alkphor での成績との相関性から, セルロースアセテート膜電気泳動では, 小腸様ALPは肝/骨ALPの位置(α2~α2β位) に比較的収束したバンドとして存在する可能性が高い. また血液型依存性高分子小腸ALPは, 骨ALPから通常の小腸ALPの位置 (α2β~β位) にかけてのブロードなバンドであると考えられた. 一方, アガロースゲル電気泳動では, 小腸様ALPおよび血液型依存性高分子小腸ALPは1本の収束したバンドとして骨ALP付近の位置 (α2β位) に検出される. したがって, 現時点においてALPアイソザイム分画の同定とその評価を行うには, 一長一短があるものの, 各種小腸ALPを比較的たやすく分別可能なPAG電気泳動を用いることが望ましいと考えられる.
  • 津村 真由美, 上野 芳人, 守尾 一昭, 桑田 昇治, 木野 内喬, 菰田 二一
    1998 年 42 巻 3 号 p. 169-174
    発行日: 1998/08/15
    公開日: 2009/03/31
    ジャーナル フリー
    われわれは, PAG電気泳動法を用い, 慢性腎不全患者の血清および尿ALPアイソザイムの解析を行ったところ, 分離ゲル上部に出現する atypical ALP-1 band を発見した. 従来進行性腎炎のマーカーとしては, NAGやβ2マイクログロブリン (β2MG) が利用されてきた. そこで, 今回はこの atypical ALP-1とβ2MGの相関性を求めた. 対象は, 糖尿病性腎症患者 (DM) と糖尿病を合併していない慢性腎不全患者 (non DM) である. 分離ゲル上部に出現する atypical ALP-1 band は, 慢性腎不全患者で高頻度に認められた. Atypical ALP-1の出現頻度は, 血清, 尿中とも non DMに比べDMで有意に高かった. DMと non DMの尿中β2MG値は, non DMに比べDMで有意に高値であった. 血清中および, 尿中の atypical ALP-1の出現はβ2 MGと密接な関係が示唆され, 腎尿細管障害を反映するものとして利用できるものと考えられた.
  • 豊田 陽子, 上野 芳人, 高階 成実, 津村 真由美, 桑田 昇治, 木野内 喬, 小澤 仁子
    1998 年 42 巻 3 号 p. 175-179
    発行日: 1998/08/15
    公開日: 2009/03/31
    ジャーナル フリー
    We detected the extra band cathodal to the electrophoretic origin through the analysis of CK isozyme of 346 healthy subjects. This band was observed in all the fresh sera stored at 4°C less than 24h. Biochemical analysis disclosed that neither adenylate kinase nor immunoglobulin-bound CK generated this band. The shift of the extracts migration after the incubation with urea implied that this extra band was derived from mitochondrial CK. We furthermore confirmed the high activation energy of this band (111.75kJ/mol) which was compatible with that of mitochondrial CK. These results strongly support that this extra band was identical with mitochondrial CK. In conclusion presumed mitochondrial CK was observed in the fresh sera of all the healthy subjects. Ordinary cell turnover may be postulated to contribute to this extra band but further study will be needed to define the precise features of this band.
  • 橋本 卯巳, 日吉 基文, 田窪 孝行, 田川 進一, 巽 典之
    1998 年 42 巻 3 号 p. 181-184
    発行日: 1998/08/15
    公開日: 2009/03/31
    ジャーナル フリー
    An Amplicor PCR series (Roche, Branchburg, NJ, USA) was developed to make the PCR technology available in hospital as a routine test, and is prevailing for the diagnosis of Mycobacterium tuberculosis, Cytomegalovirus (CMV), etc. In these circumstances, we sometimes experience a discrepancy between the result of Amplicor diagnosis and the patients' clinical findings. A method is described here to recheck the Amplicor results by performing agarose gel electrophoresis with the PCR products obtained from Amplicor. Amplicor CMV PCR test was used for this study. When the PCR products were electrophoresed without any purification, bands of amplified CMV DNA were not electrophoresed precisely. On the other hand, when the products were purified by the Qiagen blood kit (Hiden, Germany), the PCR products were electrophoresed properly. We concluded that the best way to recheck the Amplicor results was to run electrophoresis after purification of PCR products.
  • 栗原 由利子, 横山 和則, 芝 紀代子, 亀井 幸子
    1998 年 42 巻 3 号 p. 185-188
    発行日: 1998/08/15
    公開日: 2009/03/31
    ジャーナル フリー
    We analyzed glycosaminoglycans and proteins in the exudate of an esthesioneuroblastoma. The total protein concentration was 44.0g/l. Exudate concentrations of IgG, IgA, and IgM were 6.0g/l, 2.6g/l, and 12.6g/l, respectively. IgM λ-type M-protein was found in the exudate sample. The glycosaminoglycan contents were 32.4% hyaluronic acid and 39.3% heparan sulfate. Both existed in high concentrations in the exudate.
  • 中島 元夫, 豊島 美菜子, 山形 貞子, 山形 達也
    1998 年 42 巻 3 号 p. 189-196
    発行日: 1998/08/15
    公開日: 2009/03/31
    ジャーナル フリー
    Cancer is the major mortality in most advance countries and the development of efficient diagnosis and prognosis tools for cancer therapy is one of the major goals of cancer research. Highly sensitive, reliable and convenient assays using non-invasive means are required to invent new cancer diagnostic protocols. Zymography of serum and plasma samples by electrophoresis and incubation in a substrate-embedded polyacrylamide gel is suitable for such routine diagnostic assays. Zymography is also a unique method to discover unidentified glycosidases capable of specifically cleaving a certain type of glycosaminoglycans such as hyaluronic acid and chondroitin sulfate. Here we first report the new application of zymography to detect novel glycosaminoglycanases in human malignant cancers using chemically-modified chondroitin sulfate embedded in polyacrylamide gels. Secondly, we demonstrate a correlation between plasma matrix metalloproteinase activity and tumor progression with metastasis using gelatin zymography and then discuss a possible application of zymography to cancer diagnosis.
  • 志村 清仁
    1998 年 42 巻 3 号 p. 197-202
    発行日: 1998/08/15
    公開日: 2009/03/31
    ジャーナル フリー
    The use of polyelectrolytes as mobile affinity matrices in electrophoresis has led to the development of a specific separation method for biological molecules, affinophoresis. The conjugate of a polyelectrolyte and an affinity ligand is called an affinophore. Electrophoresis of proteins in the presence of an affinophore results in a change in the mobility of a specific protein by the formation of a specific complex through the affinity of the protein to the ligand. Polylysine is useful as a base polymer of affinophores and has been used successfully as an anionic matrix after succinylation. Affinophoresis of proteases, lectins and antibodies has been carried out in agarose gel and the mobility of the proteins having affinity to each ligand was specifically changed. Two-dimensional affinophoresis, in which an affinophore was included only in the second-dimensional electrophoresis, was effective for the identification of the molecule having affinity to the affinophore in a complex mixture. The use of capillary electrophoresis highlighted the relevance of affinophoresis in quantitative analyses of molecular interactions, allowing the determination of affinity constants with small amount of samples and with high precision.
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