| JaLCDOI | 10.18926/11756 |
|---|---|
| Title Alternative | 大腸菌を用いたフォスファカン（コンドロイチン硫酸プロテオグリカン）の融合コア蛋白の発現条件の検討 |
| FullText URL | 006_063_072.pdf |
| Author | Ito, Sekiko| Okamoto, Motoi| |
| Abstract | Optimal conditions for expressing a specific region of core protein of phosphacan, a chondroitin sulfate proteoglycan known as receptor type protein tyrosine phosphatase, as fusion protein with glutathione S-transferase (GST) in E.coli were examined. DNA fragments inserted into the expression vector (pGEX-4T-1) were amplified by RT-PCR using mRNA purified from E18 rat brain as template. Primers attached with BamH I or EcoR I restriction site on 5' end were used to amplify first strand cDNA by PCR. Before ligation into the pGEX-4T-1 for GST fusion protein, PCR products were once cloned using T-A cloning system because they were not directly ligated into the pGEX-4T-1. E.coli strain BL21 was transformed by pGEX-4T-1 ligated with restriction DNA fragment cut out from pCR II plasmid vector of T-A clonig system. The growth of transformed BL21 was not different between the colony incubated at 37℃ for 24-48h and the colony stored at 4℃ for 7-10 days after 24h incubation at 37℃. The desirable OD(550) of culture medium for inducing the expression of fusion protein by isopropylthio-β-D-galactoside (IPTG) was from 0.6 to 1.0, because expression of native E.coli proteins per ml of culture medium was increased relatively when IPTG was added at OD(550) more than 1.0. The expression of fusion protein reached plateau around 6h after the induction. Relative expression of native E.coli proteins per ml of culture medium increased thereafter. Therefore, it may be desirable to purify the fusion protein around 6h after the induction. |
| Keywords | phosphacan (フォスファカン) glutathione S-transferase (グルタチオン-S-トランスフェラーゼ) BL21 IPTG fusion protein (融合蛋白) |
| Publication Title | 岡山大学医療技術短期大学部紀要 |
| Published Date | 1996-02-29 |
| Volume | volume6 |
| Start Page | 63 |
| End Page | 72 |
| ISSN | 0917-4494 |
| language | English |
| File Version | publisher |
| NAID | 120002313693 |
| JaLCDOI | 10.18926/11715 |
|---|---|
| Title Alternative | An experimental study on the relation of T2-signal high intensity in MRI to histopathological changes in the kainic acid model of temporal lobe epilepsy in rats. |
| FullText URL | 010_2_069_076.pdf |
| Author | Sakiyama, Junko| Okamoto, Motoi| Kitamura, Yoshihiro| Yamada, Norihito| |
| Abstract | 側頭葉てんかんでは，てんかん焦点に一致してMRI T2高信号領域が見られ，FLAIR法でこれがより明瞭になるが，このMRI所見と病理組織学的変化との関係は必ずしもはっきりしていない。そこで，Sprague－Dawleyラットにカイニン酸（KA）でけいれん発作重積状態を起こし，経時的にMRIを記録するとともに，ニッスル染色，GFAP免疫染色での病理組織学的変化を調べて両者の関係について検討した。KA群では，MRIで1～8週間後のいずれにおいてもpiriform cortexからentorhinal cortexにかけて不整形のT2高信号領域がみられたが，stage3のけいれん発作しか出現しなかったラットではstage4，5が出現したラットに比べて程度が弱かった。組織学的には，CA1，subiculum，piriform cortex，entorhinal cortexで神経細胞の消失，濃染細胞の増加と萎縮，GFAP免疫反応の増強が見られたが，piriform cortex，entorhinal cortexでの神経細胞消失の程度はT2信 号の程度と相関せず，GFAP免疫反応が増強した領域に一致して高信号がみられた。しかし，海馬のGFAP免疫反応増強はMRI所見に反映されず，これはMRIの解像度の限界にもよると考えられた。 |
| Keywords | カイニン酸 (kainic acid) MRI FLAIR法 (FLAIR) 神経細胞死 (neuronal death) GFAP |
| Publication Title | 岡山大学医学部保健学科紀要 |
| Published Date | 2000-03-24 |
| Volume | volume10 |
| Issue | issue2 |
| Start Page | 69 |
| End Page | 76 |
| ISSN | 1345-0948 |
| language | Japanese |
| File Version | publisher |
| NAID | 120002313709 |
| Author | 岡本 基| |
|---|---|
| Published Date | 1983-12-31 |
| Publication Title | |
| Content Type | Thesis or Dissertation |
