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題 名 | 臺灣蘇力菌CrylAc5殺蟲基因選殖及表現=Cloning and Expression of the Insecticidal Crystal Protein Gene, CrylAc5, of Bacillus Thuringiensis D4-01 from Taiwan |
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作 者 | 陳韋如; 李國欽; 高穗生; 曾耀銘; 曾經洲; | 書刊名 | 植物保護學會會刊 |
卷 期 | 48:1 民95.03 |
頁 次 | 頁17-30 |
分類號 | 433.85 |
關鍵詞 | 蘇力菌; 殺蟲基因; 基因選殖及表現; Bacillus thuringiensis; CrylAc5; Insecticidal crystal protein gene; Cloning; Expression; |
語 文 | 中文(Chinese) |
中文摘要 | 聚合酶連鎖反應(polymerase chain reaction,PCR)技術,對蘇力菌台灣分離株 D4-01 ,進行 cry 1Ac 型毒蛋白 基因的全長增幅,隨後接入載體 yT&A ,成為質體 p1AcTA1 。由質體 p1AcTA1 與蘇力菌表現載體 pSB909.5 ,構築出可表現毒蛋白的質體 p 1Ac 5SB ,並以電穿孔法送入 無質體之蘇力菌 Cry - B 中,得到轉殖的蘇力菌株 1Ac 5SB ,並可測得 130 kDa 的毒蛋白產物。 分析 此選殖之 cry 1Ac 核酸序列與其編碼之胺基酸序列,發現在基因序列上 與已知的 cry 1Ac 5 基因最相似,僅 2 個鹼基之差異,而胺基酸序列方面則完全相同。 轉殖株 1Ac 5SB 具良好的殺蟲活性,以總蛋白濃度 25 ppm(有效成份蛋白表現量 11.75 ppm) 處理擬尺蠖(Trichoplusia ni) 2 齡幼蟲, 72小時後,可得 60~ 80%的致死率。等量處理小菜蛾(Plutella xylostella) 3 齡幼蟲, 72小時後,可達80~90%的致死率。 |
英文摘要 | The rice water weevil (RWW), Lissorhoptrus oryzophilus Kuschel, was first observed in Taichung County in the second-crop rice of 2000, and soon spread to the coastal towns of Taichung County in central Taiwan in the first-crop rice of 2001. Subsequently, RWW further invaded the northwestern part of Changhua County in the first-crop rice of 2002, and the infested area reached to 8,100 ha, although the southern spreading of RWW has been retarded in subsequent years. This study investigated the inhabiting sites of adult RWWs during rice fallow periods in the winter and summer seasons to better understand their migration characteristics. In the period when rice is not planted in the winter season, adults were detected in 16.7%~33.3% of samples from rice stubble. The results indicated that adult RWWs hibernate at the base of rice stubble during fallow winter periods. Otherwise, adult RWWs also hibernate in the surface soil and leaf litter of bamboo groves, woodland groves, and at the base of grasses clumps and ground trash on levees. After harvesting the first cropping rice, adult RWWs were detected in 50%~75% of samples from rice stubble and 25%~58.3% from levees. Furthermore, during the fallow periods of the summer season, adult RWWs were not detected in the soil or leaf litter of bamboo groves or woodland groves; but fed on Gramineae grasses on levees and newly germinated rice seedlings from dropped grain in paddy fields. This indicates that newly emerged adults do not have an obvious dormant behavior at this time. The RWW has 2 generations annually in central Taiwan. The density of adult RWWs reached its peak in mid-March and larvae were observed on the root portions in late March. The maximal larval density occurred in late April. Pupae were found primarily on the root portions in early May. The first-generation adults began to pupate in late May. During the second cropping rice season, larvae were observed mostly in mid-August, with maximal densities from late August to early September. Second-generation adults began to pupate in mid-September. |
本系統中英文摘要資訊取自各篇刊載內容。