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題 名 | 香蕉轉殖細胞利用葡萄糖苷酸酶活性組織化學染色之改進=Improvement of Histochemical Staining for β-glucuronidase (GUS) Activity in Transformed Banana Cells |
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作 者 | 廖玉琬; 徐善德; 黃鵬林; | 書刊名 | 中國園藝 |
卷 期 | 45:3 1999.09[民88.09] |
頁 次 | 頁239-244 |
分類號 | 435.371 |
關鍵詞 | 香蕉; 葡萄糖苷酸酶; 組織化學染色法; 基因轉殖; Banana; β-glucuronidase; GUS; Histochemical staining; Gene transformation; |
語 文 | 中文(Chinese) |
中文摘要 | 在基因轉殖試驗中,β-葡萄糖?酸?(β-glucuronidase;GUS)基因是應用最廣泛的報導基因(reporter gene)。本試驗針對香蕉轉殖細胞及非轉殖細胞,試驗五種GUS作用基質配方,以檢視GUS組織化學活性。結果顯示,最適用於香蕉轉殖細胞的GUS組織化學染色法,乃是先將組織以含有1% Triton X-100的磷酸根之磷酸鈉系列緩衝液(50mM sodium phosphate buffer, pH 7.0)前處理,置於37℃下二小時後,再以不含Triton X-100之緩衝液清洗兩次,而後加入含有1.0mM 5-bromo-4-chloro-indoly1-β-glucuronic acid X-gluc的磷酸緩衝液及20%甲醇,可以得到最好的染色效果。 |
英文摘要 | The bacterial β-glucuronidase (GUS) gene is advantageous over other genes when introduced into plants as a reporter gene. The histochemical assay of β-glucuronidase activity, as described by Jefferson (1987), is excellent for many dicotyledonous plants. However, some researchers encountered difficulties with monocotyledons. Therefore, we examined a series of histochemical β-glucuronidase assays for transformed banana cells. The best method was that the banana calli were incubated, first, in phosphate buffer (50 mM Sodium phosphate buffer, pH 7.0) containing 1% Triton X-100 at 37℃ for 2 hours. The buffer, was then removed and fresh phosphate buffer containing 1.0 mM 5-bromo-4-chloro-indoly1-β-glucuronic acid (X-gluc) and 20% methanol was added to the calli. This method enabled us to determine β-glucuronidase gene expression more effectively for transformed banana cells. |
本系統中英文摘要資訊取自各篇刊載內容。