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題名 | 粒子槍在人參及油點百合基因轉殖之研究=Studies on Genetic Transformation of Panax Ginseng C. A. Meyer and Drimiopsis Kirkii Baker through Particle Gun Bombardment |
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作者 | 蘇宗振; 張唯勤; 邢禹依; 許華欣; Su, Tsung-jen; Chang, Wei-chin; Hsing, Yue-ie; Hsu, Wha-hsing; |
期刊 | 臺灣農業 |
出版日期 | 19970600 |
卷期 | 33:3 1997.06[民86.06] |
頁次 | 頁115-128 |
分類號 | 434.28 |
語文 | chi |
關鍵詞 | 基因轉殖; 粒子槍; GUS基因; NPTII基因; Gene transfer; Particle gun bombardment; GUS gene; β-glucuronidase gene; NPTII gene; Neomycin phosphotransferase II gene; |
中文摘要 | 本試驗利用已建立體胚發生及植株再生的人參(Panax ginseng C. A. Meyer)胚性癒合組織和油點百合(Drimiopsis kirkii Bkaer)瓶苗葉片為材料。以粒子槍法(partcle bombardment, PDS1000/He, Bio-Rad),嘗試用GUS(β-glucuronidase gene)基因為標誌基因(marker gene),轉殖入上述植物的組織中。結果發現將含GUS基因的pBI221質體DNA附著在1.6 μm的金粒上,配合粒子槍之最佳操作條件,在人參及油點百合培植體上,皆可得到 GUS 活性的短暫表現。經粒子槍轉殖後的培植體,培養在分化培養基上,可產生體胚再生成植株,並可在油點百合得到轉殖後的轉殖植株。藉由GUS活性的檢測,聚合�t連鎖反應及南方氏雜交法檢測,可測得GUS基因的表現。另以pBI121質體DNA(含GUS及NPTII基因)作為油點百合之轉殖基因,可得轉殖體胚。本試體結果顯示外來基因可藉由粒子槍的途徑達到轉殖目的。 |
英文摘要 | In the studies, embryogenic callus of Panax ginseng C.a. Meyer and leaves of Drimiopsis kirkii Baker were used for gene transformation with a marker gene (β-glucuronidase gene, GUS gene) by using particle bombardment techniques. The GUS gene contained in pBI221 (3 kb) and pBI121 (13 kb) were coated to gold particles (dimeter 1.6um) and were delivered to target explants by a setup (PDS 1000/He) of Bio-Rad. Bombarded explants. of P. ginseng C.A. Meyer and D. kirkii Baker expressed the foreign gene activity as checked with X-gluc assays after 48 hours. Transgenic plants and stable expression were found of D. kirkii Baker. Positive response of the transgenes were confirmed by GUS assay, PCR (polymerase chain reaction) , and Southern blot hybridization analysis. |
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