檢測損害或影響神經系統之化學物質樣品及篩選治療神經系統疾病之經分離之轉殖基因哺乳動物神經細胞

專利類型

發明

專利國別 (專利申請國家)

中華民國

專利申請案號

099109995

專利證號

I 401434

專利獲證名稱

檢測損害或影響神經系統之化學物質樣品及篩選治療神經系統疾病之經分離之轉殖基因哺乳動物神經細胞

專利所屬機關 (申請機關)

臺北醫學大學

獲證日期

2013/07/11

技術說明

將含有大鼠戴奧辛反應單元(Dixoin responsive element, DRE)與限制酶(restriction enzyme)DNA序列，連同含有螢火蟲冷光酶(luciferase)的載體，分別以限制酶處理；使兩者產生相同的連接位。再以T4噬菌體連接脢(ligase)將上述DNA序列與冷光酶載體接合成戴奧辛類化合物偵測再以微脂體(liposome)將偵測載體與抗藥性載體同時轉染送入大鼠神經膠質瘤(glioma)細胞後長期投予藥物，篩選出具抗藥性載體的神經膠質瘤細胞株。並萃取抗藥性細胞之染色體反應證實載體穩定鑲嵌於細胞染色體；最後對細胞株投予戴奧辛類化合物，誘發螢火蟲冷光以確認細胞株偵測污染物之靈敏度。 Rat dioxin responsive element (DRE) complementary sequences (DRECS) containing three DRE and restriction recognition sequences were synthesized. The DRECSs were cloned by inserting DNA fragment between the KpnI and NheI sites of pGL2-promoter (pGL2 pro.). The pGL2 pro/DRE was constructed by ligation of purified DRECS-KpnI/NheI and pGL2 pro-KpnI/NheI with T4 DNA ligase. The ligated plasmid was transformed into E. coli competent cells, selected by Amp-agar plate and then extracted the plasmid DNA. The obtained constructs were extracted and transfected respectively into rat C6 glioma cells by Lipofectamine™ 2000. To generate pGL2 pro/DRE stable clones, pGL2 pro/DRE and pCMV-Tag 2B were co-transfected into C6 by Lipofectamine™ 2000. These stable clones were obtained by Geneticin® (G418) resistance screening. The metabolic degradable AhR agonist, 3-methylcholanthere (3MC), was used to examine the luciferase activity of pGL2 pro/DRE and intact pGL2 C6 stable clones.

備註

本部(收文號1080069658)同意該校108年10月22日北醫校事字第1080003830號函所報終止維護專利。

連絡單位 (專責單位/部門名稱)

技術移轉中心

連絡電話

02-6638-2736-2006

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