查詢結果分析
來源資料
相關文獻
- 抗嘉磷塞基改大豆之CP4 EPSPS西方墨點法免疫分析
- 進口大豆抗嘉磷塞除草劑之測定及其基因特性之探討
- 臺灣市售抗固殺草基改大豆之篩選及其基因特性分析
- 臺灣市售飼料玉米抗嘉磷塞基因特性及檢測利用之探討
- Temperature Effect on the Sensitivity of ELISA, PA and WB to Detect Anti-HIV-1 Antibody and Infectivity of HIV-1
- 新近斷奶仔豬蛋白酶處理大豆粉的探討
- Detection of Endogenous Digitalis-Like Immunoreactive Factors in Human Blood
- The Regulatory Role of Plasma Membrane Proton-Pumping ATPase in Salt Tolerance of Soybean Plant Growing Under the Salt-Stress Condition
- 乙烯及ABA抑制劑對大豆種間雜交授粉成功率之影響
- 即食性沙拉用生菜中Escherichia coli O157:H7之污染
頁籤選單縮合
| 題 名 | 抗嘉磷塞基改大豆之CP4 EPSPS西方墨點法免疫分析=Western Blot Immunoassay of Glyphosate Tolerant Genetic Modified Soybean Based on CP4 EPSPS |
|---|---|
| 作 者 | 袁秋英; 林李昌; 謝玉貞; 蔣慕琰; | 書刊名 | 作物、環境與生物資訊 |
| 卷 期 | 3:3 民95.09 |
| 頁 次 | 頁239-248 |
| 分類號 | 434.214 |
| 關鍵詞 | 基因改造生物體; 大豆; 嘉磷塞抗藥性; 免疫分析; 西方墨點法; Genetically modified organism; GMO; Soybean; Glyphosate-resistance; CP4 EPSPS; Immunoassay; Western blot; |
| 語 文 | 中文(Chinese) |
| 中文摘要 | 摘要 本研究以自備之抗嘉璘塞 5-enolpyruvylshikimate-3-phosphate synthase (Agrobacterium sp. strain CP4, CP4 EPSPS) 抗體,利用西方墨點法(Western blot)探討 CP4 EPSPS 抗體之靈敏度及專一性,以及大 豆相關加工品之檢測。由抗嘉磷塞大豆 (Roundup Ready® Soybean, RRS)葉片抽取 genomic DNA,設計CP4 EPSPS 基因含 Hind III 及Xho I 切位之引子,進行PCR 反 應,於抗嘉磷塞大豆植材的基因組DNA 中 可增幅約380 bp 的核酸片段,非抗嘉磷塞者 則無此核酸片段。經由轉殖於大腸桿菌,選 殖含約380 bp inserted DNA 的單一菌株, 確證此增幅之核酸片段為CP4 EPSPS 第 206Met 至325Ile 胺基酸,長度為375 bp。將此 PCR 產物接合於pET-28a (+)載體,再轉殖於 大腸桿菌BL21 strain,以IPTG 誘導CP4 EPSPS 融合蛋白質表現,經純化回收單一蛋 白質(約16 KDa)製備為CP4 EPSPS 多株抗 體。以Roundup Ready®大豆標準品(0、0.1、 0.5、1、2、5 及100%)、抗嘉磷塞及非基改 之大豆葉片與種子、傳統市場及超市之豆 漿、豆腐、豆花以及豆干樣品,進行西方墨 點免疫分析。結果顯示總蛋白質含量約20 μg 之大豆標準品(0.5%以上)皆可於約47 KDa 處 呈現一明顯條帶。抗嘉磷塞大豆葉片的CP4 EPSPS 蛋白質含量為種子的3 倍以上。傳統 市場及超市未標示為非基改之豆漿、豆腐及 豆花樣品,皆可檢出CP4 EPSPS 蛋白質。本 研究自製之CP4 EPSPS 抗體,具有發展為酵 素連結免疫吸附法(Enzyme-Linked Immunosorbent Assay, ELISA)及免疫條 帶法(strip)之潛力,可應用於快速及大量篩檢 含CP4 EPSPS 轉基因作物。 |
| 英文摘要 | ABSTRACT In this study, lab-made antibody and western blot were used to investigate the sensitivity and specificity of immunoassay for glyphosate tolerant soybean and related foodproducts. Genomic DNA was extracted from glyphosate tolerant GM soybean and PCR was run using primers with Hind III and Xho 1 sites for CP4 EPSPS. The ca. 380 bp fragment was transformed to E. coli DH5α. Subsequent selection of transformed cell and sequencing of DNA showed that amplified fragment was 375 bp and was translated to 206Met to 325Ile of CP4 EPSPS. The 375 bp fragment was constructed on Pet-28a (+) vector and transformed to E. coli BL21 for further multiplication of CP4 EPSPS. The ca. 16 KDa protein was purified and used as antigen for production of polyclonal antibody. Western immunoblot assay using anti-CP4 EPSPS polyclonal antibody demonstrated that standard Roundup Ready Soybeans® at 20 μg total protein level could be detected. This lab-made polyclonal antibody was also effective for detection of seeds and living plants of Roundup Ready Soybeans® as well as processed products such as seeds, soymilk and Tufu. |
本系統中英文摘要資訊取自各篇刊載內容。